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#2200: LONG®R³IGF-I antibody (Rabbit) for RIA
Novozymes GroPep's LONG®R³IGF-I antibody for RIA is a purified antiserum, derived from rabbits immunized with recombinant LONG®R³IGF-I conjugated to ovalbumin.
Care should be taken when opening the vial and reconstituting the lyophilized antibody to avoid any losses.
Recommended handling for use in Radioimmunoassay (RIA):
For radioimmunoassay we recommend the purified antibody be reconstituted as directed below to yield a final dilution of 1/2,000. At this dilution the antibody binds 25-30% of fresh [125I]LONG®R³IGF-I tracer. Half-maximal inhibition (ED50) of radioligand binding is produced by 200 - 250 pg of LONG®R³IGF-I and the limit of detection is 10 pg / tube (or 50 pg / ml in the original sample).
Procedure:
Dissolve the vial contents in a final volume of 10ml RIA buffer (30 mM sodium phosphate pH 7.5 containing 10mM EDTA, 0.2 g/l protamine sulphate, 0.5 ml Tween-20 and 0.25 g/l sodium azide). Add 50 ul of the reconstituted antibody to each RIA tube (except the antibody-"negative" blanks) in a final incubation volume of 300 µl.
Each tube should also contain approximately 20,000 cpm of fresh [125I] LONG®R³IGF-I (80 Ci/g). Bound radioactivity is then precipitated using conventional centrifugation procedure after addition of anti-rabbit IgG, rabbit IgG and chilled PEG.
See Protocol:
#3001 Procedure for the Iodination of IGFs.
#3002 Determination of IGF-I and IGF-II in a Range of Species by Radioimmunoassay (RIA).
Results:
Anti-LONG®R³IGF-I antibody is specific for LONG®R³IGF-I, with no significant crossreactivity observed with the addition of up to 100ng of the following peptides: human IGF-I, rat IGF-I, chicken IGF-I, human IGF-II, insulin and porcine GH.
Note: bovine and porcine IGF-I were not tested since they have the same amino acid sequence as human IGF-I.
* NOT FOR USE IN HUMANS
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Thebarton SA 5031
PO Box 10065 BC
Adelaide SA AUSTRALIA
Telephone +61 (0)8 8354 7700
Facsimile +61 (0)8 8354 7788
Email:
AUAD-bioreagents@novozymes.com
Website: www.gropep.com.au
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