Result

“Some Specificity detected. However due to low antibody concentration result was inconclusive. Peptide test on the antibody is only showing weak signal.“ Q . Fang, Western University

Lane No. Antigen Loading amount Primary antibody Primary antibody dilution ratio

Secondary

antibody dilution ratio

Visualiza-tion time

1 H2BK43me3/2/1

/0 peptide 100 nM

STJ97165 H2BK43me3

antibody

1:1000 1:5000 120S

Materials for Validation: Short histone peptide

Method of validation: Peptide dot blotting

Primary Antibody: STJ97165 H2BK43me3 anti-

body

Secondary Antibody HRP goat anti-rabbit

Reference antibody: Yenzym antibody

Dilution ratio: 1:1000

Protocol

Treatment of materials: 2ul of 100nM peptide was put onto nitrocellulose membrane

Air Dry: 15 mins.

Blocking: 1X TBST with 5% skimmed milk powder for 1 hr at room temperature.

Membrane wash: 10 minutes with TBS-T

Primary antibody incubation: Primary antibody incubated for 2 hours at room temperature.

Membrane wash: 3x 10 minutes with TBS-T.

Secondary antibody incubation: secondary antibody incubated for 1 hour at room temperature.

Membrane wash: 3x 10 minutes with TBS-T

Membrane incubation: Incubate the membrane with ELC reagent for 2 minutes at room temperature.

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