InvivoGen Catalog 2011 - IBIAN Technologies fileInvivoGen offers a comprehensive list of cytokine genes classified in different families including interleukins and chemokines. PRODUCT

4CYTOKINE SIGNALING

Cytokine Genes 104

Cytokine Antibodies 104

Cytokine Reporter Cells 105-116

Cytokine and Related Genes

Description

Cytokine and related genes are provided either in a pORF or a pUNOplasmid (see page 37), both containing the complete coding sequence fromthe ATG to the Stop codon. Each gene is fully sequenced.

Gene families in pORF plasmids:- Chemokine genes - Immune receptor genes- Chemokine receptor genes - Interferon genes- Cytokine genes - Interleukin genes- Cytokine suppressor genes - Interleukin receptor genes

Gene family in pUNO plasmids:- Interferon signaling genes

Contents and Storage

Each pORF and pUNO plasmid is provided as a lyophilized transformedE. coli strain on a paper disk. Transformed strains are shipped at roomtemperature and should be stored at -20°C. Each pORF is provided with 4 pouches of E. coli Fast-Media® Amp (2 TBand 2 Agar) and each pUNO plasmid with 4 pouches of E. coliFast-Media® Blas (see pages 44-45).

Cytokines are key modulators of the immune system. They are produced mainly by T lymphocytes in response to infectious agents. Some

cytokines induce the inflammatory response or stimulate B cells to produce antibodies while others act to offset these effects by

downregulation. InvivoGen offers a comprehensive list of cytokine genes classified in different families including interleukins and chemokines.

PRODUCT QUANTITY CAT. CODE*

pORF-<gene> E. coli disk porf-<gene>

pUNO-<gene> E. coli disk puno-<gene>

* Catalog codes are available on our website

Anti-Cytokine Neutralizing IgA Antibodies


Each neutralizing IgA antibody is provided lyophilized from a 0.2 μmfiltered solution in PBS. Product should be reconstituted in sterile water.Lyophilized antibodies are stable greater than six months when storedat -20ºC. Reconstituted IgAs are stable 1 month when stored at 4ºC and6 months when aliquoted and stored at -20ºC.

Description

Neutralizing IgA antibodies are chimeric monoclonal antibodies in whichthe constant domains of the human IgA molecule were combined withmurine variable regions. They have been selected for their ability toefficiently neutralize the biological activity of selected cytokines. Theneutralizing activity of these IgA antibodies was determined usingInvivoGen’s HEK-Blue™ Cytokine Cells.

PRODUCT ANTIGEN REACTIVITY QUANTITYCATALOG

CODE

Anti-hCD40L-IgA2 Human CD40 ligand Human 100 μg maba-h40l

Anti-hIFNa-IgA2 Human Interferon a Human 100 μg maba-hifna

Anti-hIFNg-IgA2 Human Interferon g Human 100 μg maba-hifng

Anti-hIL-1b-IgA2 Human Interleukin 1b Human 100 μg maba-hil1b

Anti-hIL-4-IgA2 Human Interleukin 4 Human 100 μg maba-hil4




Anti-hTGFb-IgA2 Human Tumor Growth Factor b Human 100 μg maba-htgfb

Anti-hTNFa-IgA2 Human Tumor Necrosis Factor a Human 100 μg htnfa-mab7

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Blue™ Cytokine Reporter Cells

Blue™ Cytokine Reporter Cells is an expanding family of engineered cell lines designed to provide a simple, rapid

and reliable method to monitor the activation of signaling pathways induced by key cytokines. They allow to detect

these biologically active cytokines and can also be used to screen for compounds exhibiting agonist and antagonist

activities. Blue™ Cytokine Reporter Cells include B16-Blue™ and HEK-Blue™ Cells. B16-Blue™ and HEK-Blue™ Cells

are engineered murine melanoma and HEK293 cells, respectively. They express an inducible secreted embryonic

alkaline phosphatase (SEAP) reporter gene that can be quantitatively detected using QUANTI-Blue™, a SEAP

colorimetric detection medium. Blue™ Cytokine Reporter Cells express this SEAP reporter gene under the control

of specific promoters that are selectively activated by the cytokines or other immune modulators known to induce

the pathway of interest. Therefore, in the presence of the cytokine, agonist or antagonist compound, the pathway

is activated or inhibited modulating the SEAP activity. The amount of SEAP secreted in the cell supernatant can

be measured spectrophotometrically with QUANTI-Blue™.

JAK/STAT Pathway Smad Pathway

• B16-Blue™ IFN-a/b Cells • HEK-Blue™ TGF-b Cells

• HEK-Blue™ IFN-a/b Cells

• HEK-Blue™ IL-4/IL-13 Cells

• HEK-Blue™ IL-6 Cells

NF-kB Pathway CD40 Pathway

• HEK-Blue™ TNF-a/IL-1b Cells • HEK-Blue™ CD40L Cells

• HEK-Blue™ IL-1b Cells

• HEK-Blue™ IL-18/IL-1b Cells

• HEK-Blue™ IL-33/IL-1b Cells NEW

CYTOKINE SENSOR CELLS IFNa IFNb IFNg IL-1b IL-4 IL-6 IL-13 IL-18 IL-33 TGF-b TNF-a CD40L INFO

B16-Blue™ IFN-a/b cells +++ +++ - - - - - - - - - - p 106

HEK-Blue™ CD40L cells - - - +++ - - - - - - +++ ++++ p 115

HEK-Blue™ IFN-a/b cells +++ +++ - - - - - - - - - - p 107

HEK-Blue™ IL-1b cells - - - ++++ - - - - - - - - p 111

HEK-Blue™ IL-4/IL-13 cells +/- - - - ++ - ++ - - - - - p 108

HEK-Blue™ IL-6 cells - - - - - ++ - - - - - - p 109

HEK-Blue™ IL-18/IL-1b cells - - - +++ - - - ++++ - - - - p 112

HEK-Blue™ IL-33/IL-1b cells - - - +++ - - - - ++++ - - - p 113

HEK-Blue™ TGF-b cells - - - - - - - - - +++ - - p 114

HEK-Blue™ TNF-a/IL-1b cells - - - +++ - - - - - - ++++ - p 110

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B16-Blue™ IFN-a/b - Murine Type I IFNs Reporter Cells

➥ Monitor the JAK/STAT pathway

➥ Monitor the RIG-I/MDA-5 pathway

➥ Detection of murine IFN-a and IFN-b

Background

Interferon-alpha (IFN-a) and interferon beta (IFN-b), play an important

role in viral infections. They bind to an IFN receptor complex consisting oftwo alpha chains (IFNAR1 and IFNAR2) and recruit JAK1 and TyK2. Thesekinases phosphorylate STAT1 and STAT2 leading to the formation of theISGF3 complex. ISGF3 binds to IFN-stimulated response elements (ISRE)in the promoters of IFN-stimulated genes (ISG) to regulate theirexpression (figure 1). IFN-a and IFN-b are produced in response to viral

pathogen associated molecular patterns (PAMPs), such as viral RNA andDNA. These PAMPs are recognized by cytoplasmic pattern recognitionreceptors including the RNA helicases RIG-I or MDA5. Stimulation ofB16-Blue™ IFN-a/b cells with RIG-I/MDA-5 agonists, such as transfected

poly(I:C), triggers the secretion of IFN-a and IFN-b.

Description

B16-Blue™ IFN-a/b cells allow the detection of bioactive murine type I IFNs

by monitoring the activation of the JAK/STAT/ISGF3 pathway. They derivefrom the murine B16 melanoma cell line of C57B1/6 origin after stabletransfection with a SEAP reporter gene under the control of the IFN-a/b-

inducible ISG54 promoter. Stimulation of B16-Blue™ IFN-a/b cells with murine IFN-a or IFN-b, or

type I IFN inducers, such as tranfected poly(I:C) or poly(dA:dT), activatesthe JAK/STAT/ISGF3 pathway and triggers the subsequent production ofSEAP (figures 2&3). Levels of SEAP in the supernatant can be easilydetermined with QUANTI-Blue™, a medium that turns purple/blue in thepresence of SEAP and by reading the OD at 655 nm.

• Detection range for mIFN-a: 102 - 104 IU/ml• Detection range for mIFN-b: 102 - 104 IU/ml

B16-Blue™ IFN-a/b cells are resistant to Zeocin™


B16-Blue™ IFN-a/b cells are grown in RPMI medium, 2 mM L-glutamine,

10% FBS supplemented with 100 μg/ml Zeocin™. Each vial contains

5-7x 106cells and is supplied with 10 mg Zeocin™, 50 mg Normocin™ and

1 pouch of QUANTI-Blue™. Cells are shipped on dry ice.

Figure 2: Stimulation of B16-Blue™ IFN-a/b cells by recombinant human andmurine IFN-a and IFN-b was assessed by measuring the levels of SEAP usingQUANTI-Blue™.

PRODUCT QUANTITY CAT. CODE

B16-Blue™ IFN-a/b cells 5-7 x 106 cells bb-ifnab

Figure 1: JAK/STAT and RIG-I/MDA-5 signaling pathways

Figure 3: Response of B16-Blue™ IFN-a/b cells following their stimulation withnaked poly(I:C) or poly(:C)/LyoVec. Induction of type I IFNs was assessed bydetermining the levels of SEAP using QUANTI-Blue™.

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Related Products

Blasticidin, page 17 Anti-hIFN-a-IgA , page 104Zeocin™, page 18 QUANTI-Blue™ page 14Poly(I:C) (HMW), page 77 Poly(I:C) (HMW)/LyoVec , page 80Poly(dA:dT) Naked, page 80 Poly(dA:dT)/LyoVec, page 80

➥ Monitor the JAK/STAT/ISGF3 pathway

➥ Detection of human IFN-a and IFN-b

Background

Type I interferons, in particular interferon alpha (IFN-a) and interferon beta

(IFN-b), play a vital role in host resistance to viral infections. They signal

mainly through the JAK-STAT pathway. Following their production, IFN-a

and IFN-b bind to a common receptor (IFNAR) and recruit the Janus

kinases (JAK1 and TyK2). JAKs phosphorylate STAT1 and STAT2, whichthen dimerize and interact with IFN regulatory factor 9 (IRF9), forming acomplex named ISGF3. ISGF3 binds to IFN-stimulated response elements(ISRE) in the promoters of IFN-stimulated genes (ISG) to regulate theirexpression (figure 1).

Description

HEK-Blue™ IFN-a/b cells allow the detection of bioactive human type I

IFNs by monitoring the activation of the ISGF3 pathway. These cells weregenerated by stable transfection of HEK293 cells with the human STAT2and IRF9 genes to obtain a fully active type I IFN signaling pathway. Theother genes of the pathway (IFNAR1, IFNAR2, JAK1, TyK2 and STAT1) arenaturally expressed in sufficient amounts. The cells were further transfectedwith a SEAP reporter gene under the control of the IFN-a/b-inducible

ISG54 promoter.Stimulation of HEK-Blue™ IFN-a/b cells with human IFN-a or IFN-b

activates the JAK/STAT/ISGF3 pathway and subsequently induces theproduction of SEAP. Levels of SEAP in the supernatant can be easilydetermined with QUANTI-Blue™, a medium that turns purple/blue in thepresence of SEAP and by reading the OD at 655 nm (figure 2).Stimulation of HEK-Blue™ IFN-a/b cells with recombinant human IFN-a

can be blocked by anti-hIFN-a-IgA, a neutralizing monoclonal antibody of

the IgA isotype (see page 104).

• Detection range for hIFN-a: 5 - 104 IU/ml• Detection range for hIFN-b: 20 - 104 IU/ml

HEK-Blue™ IFN-a/b cells are resistant to blasticidin and Zeocin™.


HEK-Blue™ IFN-a/b cells are grown in standard DMEM medium, 2mM

L-glutamine,10% FBS supplemented with blasticidin (30 μg/ml) and

Zeocin™ (100 μg/ml). Cells are provided frozen in a cryotube containing

5-7 x 106 cells and supplied with 100 μl of blasticidin at 10 mg/ml, 100 μl

of Zeocin™ at 100 mg/ml, 1 ml Normocin™ at 50 mg/ml, and 1 pouch ofQUANTI-Blue™. Cells are shipped on dry ice.

HEK-Blue™ IFN-a/b Cells - Human Type I IFN Reporter Cells


HEK-Blue™ IFN-a/b cells 5-7x 106 cells hkb-ifnab

Figure 1: JAK-STAT pathway induced by type I IFNs.

Figure 2: Stimulation of HEK-Blue™ IFN-a/b cells by human IFN-a2b, IFN-b1a andIFN-g was assessed by measuring the levels of SEAP using QUANTI-Blue™.


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Related Products

Blasticidin, page 17 Anti-hIFN-a-IgA , page 104Zeocin™, page 18 QUANTI-Blue™ page 14


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HEK-Blue™ IL-4/IL-13 Cells - IL-4 and IL-13 Reporter Cells


HEK-Blue™ IL-4/IL-13 Cells 5-7 x 106 cells hkb-stat6

HEK-Blue™ IL-4/IL-13 Kit 1 kit hkb-stat6-kit

➥ Monitor the JAK/STAT-6 pathway

➥ Detection of human IL-4 and human/murine IL-13

Background

The transcription factor STAT6 is activated primarily by two cytokineswith overlapping biologic functions, IL-4 and IL-13. It can also be activatedby IFN-a in a cell-specific manner. In non-hematopoietic cells, IL-4 and

IL-13 bind a receptor complex composed of the IL-4Ralpha andIL-13Ralpha1. Upon ligand binding, the receptor complex activates thereceptor-associated Janus kinases (JAK1 and Tyk2) leading to therecruitment of STAT6 and its phosphorylation. Activated STAT6 formshomodimers that translocate to the nucleus where they bind thepromoter of responsive genes inducing gene transcription.

Description

HEK-Blue™ IL-4/IL-13 cells allow the detection of bioactive IL-4 andIL-13 by monitoring the activation of the STAT-6 pathway. These cellswere generated by stable transfection of HEK293 cells with the humanSTAT6 gene to obtain a fully active STAT6 pathway. The other genes ofthe pathway are naturally expressed in sufficient amounts. The cells werefurther transfected with a SEAP reporter gene under the control of theIFNb minimal promoter fused to four STAT6 binding sites. HEK-Blue™

IL-4/IL-13 cells produce SEAP in response to IL-4 or IL-13 stimulationand to a lower extent IFN-a. The levels of SEAP secreted in the

supernatant can be easily determined with QUANTI-Blue™, a SEAPdetection medium (Figure 2).Stimulation of HEK-Blue™ IL-4/IL-13 cells with IL-4 or IL-13 can be blockedby the neutralizing monoclonal anti-hIL-4-IgA and anti-hIL-13-IgAantibodies, respectively (see page 104).

• Detection range for IL-4: 0.5 - 100 ng/ml• Detection range for IL-13: 5 - 1000 ng/ml

HEK-Blue™ IL-4/IL-13 cells are resistant to blasticidin and Zeocin™.


HEK-Blue™ IL-4/IL-13 cells are grown in standard DMEM medium, 2 mML-glutamine with 10% FBS supplemented with blasticidin (10 μg/ml) and

Zeocin™ (100 μg/ml). Each vial contains 5-7 x 106 cells and is supplied

with 100 μl of blasticidin at 10 mg/ml, 100 μl of Zeocin™ at 100 mg/ml,

1 ml Normocin™ at 50 mg/ml and 1 pouch of QUANTI-Blue™. Cells areshipped on dry ice.

HEK-Blue™ IL-4/IL-13 cells are also available in a kit. See below:

Figure 1: JAK/STAT6 signaling pathway induced by IL-4 andIL-13.

Figure 2: Stimulation of HEK-Blue™ IL-4/IL-13 cells by recombinant human IL-4 andIL-13 was assessed by measuring the levels of SEAP secreted in the supernatant usingQUANTI-Blue™.

HEK-Blue™ IL-4/IL-13 Kit

• HEK-Blue™ IL-4/IL-13 Cells 5-7 x 106 cells

• Anti-hIL-4-IgA antibody 100 μg


• QUANTI-Blue™ 1 pouch (100 ml)

• Normocin™ 1 ml (50 mg/ml)

• Blasticidin 100 μl (10 mg/ml)

• Zeocin™ 100 μl (100 mg/ml)

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➥ Monitor the JAK/STAT-3 pathway

➥ Detection of human IL-6

Background

The proinflammatory cytokine IL-6 is one of the most importantmediators of fever and of the acute phase response. IL-6 exerts itsaction by first binding to the IL-6R. The complex of IL-6 and IL-6Rassociates with the signal-transducing membrane protein gp130, therebyinducing its dimerization. This leads to the activation by phosphorylationof the tyrosine kinases of the Janus family (JAK1, JAK2, and Tyk2).Activated JAKs induce the dimerization and translocation to the nucleusof STAT3 where it binds enhancer elements of IL-6-inducible genes.

Description

HEK-Blue™ IL-6 cells allow the detection of bioactive IL-6 bymonitoring the activation of the STAT-3 pathway. These cells weregenerated by stable transfection of HEK293 cells with the humanIL-6R gene. They were further transfected with a SEAP reporter geneunder the control of the IFN-b minimal promoter fused to four STAT3

binding sites. Upon IL-6 stimulation, HEK-Blue™ IL-6 cells trigger theactivation of STAT3 and the subsequent secretion of SEAP. SEAP canbe readily monitored when using the SEAP detection mediumQUANTI-Blue™.Stimulation of HEK-Blue™ IL-6 cells with recombinant human IL-6 can beblocked by anti-hIL-6-IgA, a neutralizing monoclonal antibody of the IgAisotype (see page 104).

• Detection range for IL-6: 0.5 - 50 ng/ml

HEK-Blue™/IL-6 cells are resistant to both hygromycin and Zeocin™.


HEK-Blue™ IL-6 cells are grown in standard DMEM medium, 2 mML-glutamine, 10% FBS supplemented with 200 µg/ml HygroGold(ultrapure hygromycin) and 100 µg/ml Zeocin™. Cells are provided frozenin a cryotube containing 5-7 x 106 cells and supplied with 100 µl ofHygroGold™ at 100 mg/ml, 100 µl of Zeocin™ at 100 mg/ml, 1 mlNormocin™ at 50 mg/ml and 1 pouch of QUANTI-Blue™. Cells areshipped on dry ice.

HEK-Blue™ IL-6 cells are also available in a kit. See below:

HEK-Blue™ IL-6 Cells - IL6 Reporter Cells


HEK-Blue™ IL-6 Cells 5-7 x 106 cells hkb-il6

HEK-Blue™ IL-6 Kit 1 kit hkb-il6-kit

Figure 1: JAK/STAT3 signaling pathway.

Figure 2: Stimulation of HEK-Blue™ IL-6 cells by recombinant human IL-6 was assessedby measuring the levels of SEAP using QUANTI-Blue™.

HEK-Blue™ IL-6 Kit

• HEK-Blue™ IL-6 Cells 5-7 x 106 cells




• HygroGold™ 100 μl (100 mg/ml)



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HEK-Blue™ TNF-a/IL-1b Cells - TNF-a and IL-1b Reporter Cells


HEK-Blue™ TNFa/IL-1b Cells 5-7 x 106 cells hkb-tnfil1

HEK-Blue™ TNF-a/IL-1b Kit 1 kit hkb-tnfil1-kit

➥ Monitor the TNF-a- and IL-1b-induced NF-kB and AP-1 pathways

➥ Detection of human and murine TNF-a and IL-1b(a)

Background

Tumor necrosis factor alpha (TNF-a) and interleukin 1 beta (IL-1b) are

proinflammatory cytokines produced in response to microbial infection.The common inflammatory responses of TNF-a and IL-1b are mediated

by interactions of these cytokines with distinct receptors: TNF-a binds

TNFR1 and TNFR2, IL-1b binds IL-1RI. Binding of these cytokines to their

receptors induces intracellular signaling. TNF-a signaling involves TRADD,

TRAF2 and RIP, while IL-1b signaling is mediated by MyD88, IRAK-1,

TRAF-6, and Rac1. Both signalings lead to the activation of thetranscription factors NF-kB and AP-1 (figure 1).

Description

HEK-Blue™TNF-a/IL-1b cells are designed to detect bioactive TNF-a and

IL-1b (and IL-1a) by monitoring the activation of the NF-kB pathway. HEK-Blue™ TNF-a/IL-1b cells derive from HEK293 cells. They

endogenously express the receptors for TNF-a and IL-1b and stably

express a SEAP reporter gene under the control of the IFN-b minimal

promoter fused to five NF-kB and five AP-1 binding sites.

HEK-Blue™ TNF-a/IL-1b cells secrete SEAP upon stimulation by human

TNF-a and IL-1b (figure 2). SEAP production is also detected in the

presence of murine TNF-a and IL-1b, and human or murine IL-1a. SEAP

levels can be readily determined using a SEAP detection medium, suchas QUANTI-Blue™ or HEK-Blue™ Detection medium.Stimulation of HEK-Blue™ TNF-a/IL-1b cells with recombinant human

TNF-a or IL-1b can be blocked by the neutralizing monoclonal anti-

hTNF-a-IgA or anti-hIL-1b-IgA antibody, respectively (see page 104).

• Detection range for TNF-a: 0.5 ng - 1 μg/ml• Detection range for IL-1b: 0.2 - 100 ng/ml

HEK-Blue™ TNF-a/IL-1b cells are resistant to Zeocin™.


HEK-Blue™ TNF-a/IL-1b cells are grown in DMEM medium, 2 mM

L-glutamine, 10% FBS supplemented with Zeocin™ (100 μg/ml). Each vial

contains 5-7 x 106 cells and supplied with 100 μl of Zeocin™ at 100

mg/ml, 1 ml Normocin™ at 50 mg/ml and 1 pouch of QUANTI-Blue™.Cells are shipped on dry ice.

HEK-Blue™ TNF-a/IL-1b cells are also available in a kit. See below:

Figure 1: NF-kB/AP-1 signaling pathways induced by TNF-aand IL-1b.

Figure 2: Stimulation of HEK-Blue™ TNF-a/IL-1b cells by recombinant humanTNF-a and IL-1b was assessed by measuring the levels of SEAP secreted in thesupernatant using QUANTI-Blue™.

HEK-Blue™ TNF-a/IL-1b Kit

• HEK-Blue™ TNF-a/IL-1b Cells 5-7 x 106 cells

• Anti-hTNF-a-IgA antibody 100 μg

• Anti-hIL-1b-IgA antibody 100 μg




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➥ Monitor the IL-1b-induced NF-kB and AP-1 pathways

➥ Detection of human and murine IL-1b(a)

Background

Interleukin-1b (IL-1b) is an important mediator of the inflammatory response

to infection and injury. IL-1b binds to the type 1 IL-1 receptor (IL-1R) which

requires the IL-1 receptor accessory protein (IL-1RAcP) to transduce asignal. IL-1b signaling is mediated by MyD88, IRAK-1/2 and TRAF-6 leading

to the activation of NF-kB and AP-1 (figure 1).

Description

HEK-Blue™ IL-1b cells allow to detect bioactive IL-1b by monitoring the

activation of the NF-kB and AP-1 pathways. They derive from HEK-Blue™

TNF-a/IL-1b cells in which the TNF-a response has been blocked.

Therefore, HEK-Blue™ IL-1b cells respond specifically to IL-1b. They express

a SEAP reporter gene under the control of the IFN-b minimal promoter

fused to five NF-kB and five AP-1 binding sites.

Binding of IL-1b to its receptor on the surface of HEK-Blue™ IL-1b cells

triggers the IL1-R signaling pathway leading to the activation of NF-kB/

AP-1 and the subsequent production of SEAP. The levels of SEAP in thesupernatant can be easily monitored using QUANTI-Blue™.HEK-Blue™ IL-1b cells respond to IL-1b and also IL-1a (data not shown)

but do not respond to TNF-a (figure 2).

Stimulation of HEK-Blue™ IL-1b cells with recombinant human IL-1b can

be blocked by the neutralizing monoclonal anti-hIL-1b-IgA antibody (see

page 104),

• Detection range for human IL-1b: 100 pg - 100 ng/ml• Detection range for murine IL-1b: 10 ng - 1 μg/ml

HEK-Blue™ IL-1b cells are resistant to Zeocin™ and hygromycin B.


HEK-Blue™ IL-1b cells are grown in DMEM medium, 2 mM L-glutamine, 10%

FBS supplemented with 100 μg/ml Zeocin™ and 200 μg/ml HygroGold™

(ultrapure Hygromycin). Each vial contains 5-7 x 106

cells and is suppliedwith 10 mg Zeocin™, 10 mg HygroGold™, 50 mg Normocin™ and 1 pouchof QUANTI-Blue™ Cells are shipped on dry ice.

HEK-Blue™ IL-1b cells are also available in a kit. See below:

HEK-Blue™ IL-1b Cells - IL-1b Reporter Cells


HEK-Blue™ IL-1b Cells 5-7 x 106 cells hkb-il1b

HEK-Blue™ IL-1b Kit 1 kit hkb-il1b-kit

Figure 1: IL-1b-induced NF-kB signaling pathway.

Figure 2: Stimulation of HEK-Blue™ IL-1b cells by human and murine IL-1b andhuman TNF-a was assessed by measuring the levels of SEAP using QUANTI-Blue™.

HEK-Blue™ IL-1b Kit

• HEK-Blue™ IL-1b Cells 5-7 x 106 cells






HEK-Blue™ IL-18/IL-1b Cells - IL-18/IL-1b Reporter Cells


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HEK-Blue™ IL-18/IL-1bCells 5-7 x 106 cells hkb-il18

HEK-Blue™ IL-18/IL-1b Kit 1 kit hkb-il18-kit

Figure 2: Stimulation of HEK-Blue™ IL-18/IL-1b cells by human IL-18, IL-1b andTNF-a was assessed by measuring the levels of SEAP using QUANTI-Blue™.

Figure 1: IL-18- and IL-1b--induced signaling pathways.

➥ Monitor the IL-18/IL-1b-induced NF-kB and AP-1 pathways

➥ Detection of IL-18 and IL-1b(a)

Background

Interleukin-1b (IL-1b) and IL-18 are related pro-inflammatory cytokines

that cause a wide variety of biological efffects associated with infection,inflammation and autoimmune processes. IL-1b binds to the type 1 IL-1

receptor which requires the IL-1 receptor accessory protein (IL-1RAcP)to transduce a signal. IL-18 binds to an heterodimeric receptor consistingof IL-18R and IL-18 receptor accessory protein (IL18RAP). IL-1b and IL-

18 share common signaling pathways that involve MyD88, and TRAF-6leading to the activation of NF-kB and AP-1 (figure 1).

Description

HEK-Blue™ IL-18/IL-1b cells are designed to detect bioactive IL-18 and

IL-1b (and IL-1a) by monitoring the activation of the NF-kB and AP-1pathways. They were generated by stable transfection of HEK-Blue™

IL-1b cells with the gene encoding IL-18RAP. They express a SEAP

reporter gene under the control of the IFN-b minimal promoter fused

to five NF-kB and five AP-1 binding sites.

Stimulation of HEK-Blue™ IL-18/IL-1b cells with human IL-18 or IL-1b,

but not with human TNF-a, activates the NF-kB and AP-1 pathways

triggering the production of SEAP (figure 2). Levels of SEAP in thesupernatant can be easily determined with QUANTI-Blue™.Stimulation of HEK-Blue™ IL-18/IL-1b cells with recombinant human

IL-18 or IL-1b can be blocked by the neutralizing antibodies

anti-hIL-18-IgA and anti-hIL-1b-IgA, respectively (see page 104).

• Detection range for human IL-18: 0.5 - 100 ng/ml• Detection range for human IL-1b: 0.5 - 100 ng/ml

HEK-Blue™ IL-18/IL-1b cells are resistant to blasticidin, Zeocin™ and

hygromycin B.


HEK-Blue™ IL-18/IL-1b cells are grown in DMEM medium, 2mM

L-glutamine, 10% FBS supplemented with 30 μg/ml blasticidin, 100 μg/ml

Zeocin™ and 200 μg/ml HygroGold™ (ultrapure Hygromycin). Each vial

contains 5-7 x 106cells and is supplied with 1 mg blasticidin, 10 mg Zeocin™,

10 mg HygroGold™, 50 mg Normocin™ and 1 pouch of QUANTI-Blue™

Cells are shipped on dry ice. HEK-Blue™ IL-18/IL-1b cells are also available in a kit, see below:

HEK-Blue™ IL-18/IL-1b Kit

• HEK-Blue™ IL-18/IL-1b Cells 5-7 x 106 cells








HEK-Blue™ IL-33/IL-1b Cells - IL-33/IL-1b Reporter Cells NEW


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Related Products

Anti-hIL-1b-IgA , page 104 Blasticidin, page 17Normocin™, page 11 HygroGold™, page 18QUANTI-Blue™, page 14 Zeocin™, page 18 PRODUCT QUANTITY CAT. CODE

HEK-Blue™ IL-33/IL-1bCells 5-7 x 106 cells hkb-il33

➥ Monitor the IL-33/IL-1b-induced NF-kB and AP-1 pathways

➥ Detection of IL-33 and IL-1b(a)

Background

Interleukin-33 (IL-33) is a member of the IL-1 family, a group of cytokinesthat play important roles in host defense, immune regulation andinflammation. IL-33 mediates its biological effects through ST2 (also knownas IL1RL1), a receptor expressed on Th2 and mast cells. IL-33 and ST2 forma complex with IL-1R accessory protein (IL-1RAcP), a signaling receptorsubunit that is also a member of the IL-1R complex. IL-33 signaling leads tothe activation of NF-kB and MAP kinases, and the production of Th2-

associated cytokines.

Description

HEK-Blue™ IL-33/IL-1b cells are designed to detect bioactive IL-33 and

IL-1b (and IL-1a) by monitoring the activation of the NF-kB and AP-1pathways. They were generated by stable transfection of HEK-Blue™

IL-1b cells with the IL1RL1 gene. They express a SEAP reporter gene

under the control of the IFN-b minimal promoter fused to five NF-kB

and five AP-1 binding sites. Stimulation of HEK-Blue™ IL-33/IL-1b cells with human IL-33 or IL-1b,

but not with IL-18 nor TNF-a, activates the NF-kB and AP-1 pathways

triggering the production of SEAP (figure 2). Levels of SEAP in thesupernatant can be easily determined with QUANTI-Blue™.Stimulation of HEK-Blue™ IL-33/IL-1b cells with recombinant human

IL-1b can be blocked by using the neutralizing antibody anti-hIL-1b-IgA.

• Detection range for human IL-33: 0.5 - 100 ng/ml• Detection range for human IL-1b: 0.5 - 100 ng/ml

HEK-Blue™ IL-33/IL-1b cells are resistant to blasticidin, Zeocin™ and

hygromycin B.


HEK-Blue™ IL-33/IL-1b cells are grown in DMEM medium, 2mM

L-glutamine, 10% FBS supplemented with 30 μg/ml blasticidin, 100 μg/ml

Zeocin™ and 200 μg/ml HygroGold™ (ultrapure Hygromycin). Each vial

contains 5-7 x 106cells and is supplied with 1 mg blasticidin, 10 mg Zeocin™,

10 mg HygroGold™, 50 mg Normocin™ and 1 pouch of QUANTI-Blue™

Cells are shipped on dry ice.

Figure 1: IL-33- and IL-1b--induced signaling pathways.

Figure 2: Stimulation of HEK-Blue™ IL-33/IL-1b cells by human IL-33, IL-1b, IL-18and TNF-a was assessed by measuring the levels of SEAP using QUANTI-Blue™.


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HEK-Blue™ TGF-b Cells - TGF-b Reporter Cells

➥ Monitor the TGF-b/Smad pathway

➥ Detection of human TGF-b

Background

Tumor growth factor-beta (TGF-b) belongs to a family of structurally

related cytokines that regulate a plethora of cellular functions, such asproliferation, apoptosis, differentiation and migration. TGF-b binds to a

type II receptor which recruits and activates a type I receptor. The type Ireceptor then phosphorylates receptor-regulated Smads (R-Smads), suchas Smad2 and Smad3, which associate with Smad4. R-Smad/Smad4complexes accumulate in the nucleus where they regulate the

transcription of target genes.

Description

HEK-Blue™ TGF-b cells allow the detection of bioactive TGF-b by

monitoring the activation of the TGF-b/Smad pathway. They were

generated by stable transfection of HEK293 cells with the human TGFBRI,Smad3 and Smad4 genes. They further express a SEAP reporter geneunder the control of the b-globin minimal promoter fused to three

Smad3/4-binding elements (SBE).Stimulation of HEK-Blue™ TGF-b cells with TGF-b induces the activation

of the TGF-b/Smad signaling pathway leading to the formation of a

Smad3/Smad4 complex. The heterocomplex enters the nucleus and bindsSBE sites inducing the production of SEAP (figure 1). The quantity of SEAPsecreted in the supernatant can be easily assessed using QUANTI-Blue™

(figure 2). TGF-b-mediated SEAP production can be blocked using a

neutralizing antibody, such as anti-hTGF-b-IgA (see page 104).

• Detection range for TGF-b: 0.1 - 10 ng/ml

HEK-Blue™ TGF-b cells are resistant to blasticidin, hygromycin and

Zeocin™.


HEK-Blue™ TGF-b cells are grown in DMEM medium, 2 mM L-glutamine,

10% FBS supplemented with 30 μg/ml blasticidin, 200 µg/ml HygroGold™

(ultrapure hygromycin) and 100 µg/ml Zeocin™. Cells are provided frozenin a cryotube containing 5-7 x 106 cells and supplied with 100 µl ofblasticidin at 10 mg/ml, 100 µl of HygroGold™ at 100 mg/ml, 100 µl ofZeocin™ at 100 mg/ml and 1 pouch of QUANTI-Blue™. Cells are shippedon dry ice.HEK-Blue™ TGF-b cells are also available in a kit, see below:


HEK-Blue™ TGF-b Cells 5-7 x 106 cells hkb-tgfb

HEK-Blue™ TGF-b Kit 1 kit hkb-tgfb-kit

Figure 1: TGF-b-induced Smad signaling pathway

Figure 2: Stimulation of HEK-Blue™ TGF-b cells by human TGF-b was assessed bymeasuring the levels of SEAP using QUANTI-Blue™.

HEK-Blue™ CD40L Kit

• HEK-Blue™ TGF-b Cells 5-7 x 106 cells

• Anti-hTGF-b-IgA antibody 100 μg







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HEK-Blue™ CD40L Cells - CD40L Reporter Cells


HEK-Blue™ CD40L Cells 5-7 x 106 cells hkb-cd40

HEK-Blue™ CD40L Kit 1 kit hkb-cd40-kit

➥ Study CD40L-CD40 interactions

➥ Screen for molecules that interfere with CD40L-CD40 cross-talk

Background

CD40 ligand (CD40L) is a member of the tumor necrosis factor (TNF)family of cell surface interaction molecules. It is mainly expressed inCD4+-T cells and interacts with CD40 on antigen-presenting cells toregulate both humoral and cellular immune responses. CD40L-CD40interactions are thought to play an important role in the pathogenesis ofcertain diseases, including AIDS, Alzheimer’s disease and rheumatoidarthritis.

Description

HEK-Blue™ CD40L cells can serve to measure the bioactivity of CD40Lthrough the secretion of embryonic alkaline phosphatase (SEAP) uponNF-kB activation. These cells were generated by stable transfection of

HEK293 cells with the human CD40 gene and an NF-kB-inducible SEAP

construct. The SEAP construct consists of the SEAP reporter gene underthe control of the IFN-b minimal promoter fused to five NF-kB binding

sites. Binding of CD40L to its receptor CD40 triggers a signaling cascadeleading to the activation of NF-kB and the subsequent production of

SEAP (figure 1). CD40L-CD40 interaction can be monitored by assessingthe levels of SEAP using a SEAP detection assay, such as QUANTI-Blue™

(figure 2). HEK293 cells express endogenously the receptors for thecytokines IL-1b and TNF-a which share a common signaling pathway with

CD40L. Consequently, HEK-Blue™ CD40L cells also respond to IL-1b and

TNF-a. IL-1b- and TNF-a-mediated SEAP production can be blocked

using neutralizing antibodies, such as anti-hIL-1b-IgA and anti-hTNF-a-IgA

antibodies respectively (see page 104).

• Detection range for CD40L: 5 ng - 1 μg/ml

HEK-Blue™ CD40L cells are resistant to the selective antibiotics blasticidinand Zeocin™.


HEK-Blue™ CD40L cells are grown in DMEM medium, 2mM L-glutamine,10% FBS supplemented with 100 μg/ml Zeocin™ and 30 μg/ml blasticidin.

Each vial contains 5-7 x 106

cells and is supplied with 10 mg Zeocin™,1 mg blasticidin, 50 mg Normocin™ and 1 pouch of QUANTI-Blue™. Cellsare shipped on dry ice. HEK-Blue™ CD40L cells are also available in a kit, see below: Figure 2: Stimulation of HEK-Blue™ CD40L cells by human CD40L was assessed by

measuring the levels of SEAP using QUANTI-Blue™.

Figure 1: CD40L-induced NF-kB signaling pathway.

HEK-Blue™ CD40L Kit

• HEK-Blue™ CD40L Cells 5-7 x 106 cells

• Anti-hCD40L-IgA antibody 100 μg





PRODUCT CONTENTS CAT. CODE

HEK-Blue™ IL-4/IL-13 Kit • HEK-Blue™ IL-4/IL-13 Cells (5-7 x 106 cells) • Normocin™ (1 ml @ 50 mg/ml)

• Anti-hIL-4-IgA neutralizing antibody (100 μg) • Blasticidin (100 μl @ 10 mg/ml)

• Anti-hIL-13-IgA neutralizing antibody (100 μg) • Zeocin™ (100 μl @ 100 mg/ml)

• QUANTI-Blue™ (1 pouch, 100 ml)

hkb-stat6-kit

HEK-Blue™ IL-6 Kit • HEK-Blue™ IL-6 Cells (5-7 x 106 cells) • Normocin™ (1 ml @ 50 mg/ml)

• Anti-hIL-6-IgA neutralizing antibody (100 μg) • HygroGold™ (100 μl @ 100 mg/ml)

• QUANTI-Blue™ (1 pouch, 100 ml) • Zeocin™ (100 μl @ 100 mg/ml)

hkb-il6-kit

HEK-Blue™ TNF-a/IL-1b Kit • HEK-Blue™ TNF-a/IL-1b Cells (5-7 x 106 cells) • QUANTI-Blue™ (1 pouch, 100 ml)

• Anti-hTNF-a-IgA neutralizing antibody (100 μg) • Normocin™ (1 ml @ 50 mg/ml)

• Anti-hIL-1b-IgA neutralizing antibody (100 μg) • Zeocin™ (100 μl @ 100 mg/ml)

hkb-tnfil1-kit

HEK-Blue™ IL-1b Kit • HEK-Blue™ IL-1b Cells (5-7 x 106 cells) • Normocin™ (1 ml @ 50 mg/ml)

• Anti-hIL-1b-IgA neutralizing antibody (100 μg) • HygroGold™ (100 μl @ 100 mg/ml)


hkb-il1b-kit

HEK-Blue™ IL-18/IL-1b Kit • HEK-Blue™ IL-18/IL-1b Cells (5-7 x 106 cells) • Normocin™ (1 ml @ 50 mg/ml)

• Anti-hIL-18-IgA neutralizing antibody (100 μg) • Blasticidin (100 μl @ 10 mg/ml)

• Anti-hIL-1b-IgA neutralizing antibody (100 μg) • HygroGold™ (100 μl @ 100 mg/ml)


hkb-il18-kit

HEK-Blue™ TGF-b Kit • HEK-Blue™ TGF-b Cells (5-7 x 106 cells) • Blasticidin (100 μl @ 10 mg/ml)

• Anti-hTGF-b-IgA neutralizing antibody (100 μg) • HygroGold™ (100 μl @ 100 mg/ml)


• Normocin™ (1 ml @ 50 mg/ml)

hkb-tgfb-kit

HEK-Blue™ CD40L Kit • HEK-Blue™ CD40L Cells (5-7 x 106 cells) • Normocin™ (1 ml @ 50 mg/ml)

• Anti-hCD40L-IgA neutralizing antibody (100 μg) • Blasticidin (100 μl @ 10 mg/ml)


hkb-cd40-kit

116 www.inv ivogen.com/cytokine-sensor-ce l l s

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HEK-Blue™ Cytokine Kits


HEK-Blue™ Cytokine cells are shipped on dry ice. Thaw immediately or store in liquid nitrogen. Other products are shipped at room temperature and shouldbe stored at -20°C.

Documents

InvivoGen Catalog 2011 - IBIAN Technologies fileInvivoGen offers a comprehensive list of cytokine genes classified in different families including interleukins and chemokines. PRODUCT