not sure if this procedure is correct by any means..bits and pieces from different sources
Citation preview
General assay procedure for testosterone.
1. Sampling procedureA) Volume varies measured2. Recovery
corrections3. Extraction and evaporationa) Solvent Diethyl ether of
the highest grade from freshly opened can. 500ml cans. b) extract
once with 5ml etherc) vortex in a glass tube, allow to settle and
remove supernatant into clean glass tubed) place the rack of tubes
in solid CO2 (dry ice) and acetone. This is to freeze. Will allow
the supernatant to decanted. e) can also be transferred using a
Pasteur pipette.f) can be evaporated using three ways: 1-evaporate
in the fume chamber at room temp until the ether has evaporated,
2-blown using nitrogen or compressed air and they must be
filtered.3- tubes can be vacuum driedg) perform an ether blank.
(5ml) value obtained should be 95% confidence limits of the Blank
value. 4. redissolving and aliquotinga) 2ml of assay buffer should
be added. b) vortex mix, 5 minutes gap, vortex again.c) take 0.5ml
of the redissolved extract into a scintillation vial and add 10ml
of a suitable scintillation fluid using a zippette.d) duplicate
0.5ml of the aliquots are transferred to the assay tubes.e) for
each sample and standard in duplicate, add 100ul working solution,
100ul antiserum, d) addition of charcoal suspension 0.2ml
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1. Reagents of Peptide Hormones 5g of Sodium dihydrogen
Phosphate (anhydrous) (MW120) 15g of Disodium hydrogen phosphate
(anhydrous) (MW142)Reagents to make theAssay buffer.
10g of Sodium Chloride 2gs of Thimersal 10g of Bovine Serum
Albumin
2. Peptide Standards sold in lyophilized form in ampoules
$236
