Specificity of the antiserum is defined as the ratio of antigen concentration to cross-reactant concentration at 50% inhibition of maximum binding. The cross-reactivity data obtained in the described RIA system is as follows:
5 alpha-Pregnane-3,20-dione 1.8%
3 beta-Hydroxy-5-pregnane-20-one 22.2%
17 beta-Estradiol <0.1%
11 alpha-Hydroxyprogesterone 15.9%
11 beta-Hydroxyprogesterone <0.1%
20 alpha-Hydroxy-4-pregnane-3-one 0.1%
20 beta-Hydroxy-4-pregnane-3-one 4.6%
Each vial contains no more than 20 mg Polyvinylpyrrolidone (PVP).
Lyophilised:1. Stock Solution: To one vial of lyophilized powder add 5.0 ml of 0.05 M Tris-HCl buffer, pH 8.0, containing 0.1 M NaCl, 0.1% gelatin and 0.1% sodium azide. Rotate vial gently until powder is dissolved.
2. Working Solution: To obtain the number of tests indicated on the vial further dilute the reconstituted antiserum 10-fold with the buffer used to prepare the stock solution.Discard if unused within 12 hours.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Constituents: Whole Serum
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The product is provided as a pre-diluted antiserum that has been lyophilized.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
RIA: The antiserum is characterized utilizing the following dextran coated charcoal radioimmunoassay (RIA) protocol, where 0.5 ml of reconstituted and diluted antiserum has been found to bind at least 40% of 10-15 picograms of tritiated (3H) progesterone with a specific activity of approximately 100 Ci/mmole.
(A) Standards: Prepare a stock standard solution of 1 µg/ml progesterone in absolute ethanol. Dilute a portion of the stock solution with buffer (B) to a concentration of 500 pg/0.1ml. This is further diluted in buffer (B) to obtain standard solutions at the following concentrations: 250, 125, 63, 31 and 15 pg/0.1 ml.
(B) Dilution buffer: 0.05 M Tris-HCl, pH 8.0, containing 0.1 M NaCl, 0.1% BSA and 0.1% sodium azide.
(C) Dextran coated charcoal suspension: 0.5% activated charcoal untreated powder 100-400 mesh, 0.5% dextran approximate average molecular weight 70,000 in buffer (B). It is important that the dextran be in solution before the addition of charcoal. The dextran coated charcoal suspension should be stirred and kept at 0°C in ice-water for at least 30 minutes before and during use.
1. In polypropylene test tubes add 0.1 ml sample or standard (A) and 0.5 ml diluted antiserum.
2. Vortex the tubes.
3. Incubate for 30 minutes at room temperature.
4. Add 0.1 ml tritiated radioactive tracer diluted in dilution buffer (B).
5. Vortex the tubes.
6. Incubate for 1 hour at 37°C.
7. Cool the tubes for 15 minutes at 4°C.
8. Rapidly add 0.2 ml cold dextran coated charcoal suspension (C) to each tube.
9. Vortex the tubes.
10. Incubate for 10 minutes at 0°C in ice-water.
11. Centrifuge at 2000 x g for 15 minutes at 4°C.
Sensitivity is defined as the 90% intercept of a B/B0 standard curve. In the above system the sensitivity has been found to be 5pg progesterone/tube.
The affinity constant (K ) is determined by a Scatchard a plot using the described RIA system. K = 2.9 x 10 L/mole.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Progesterone plays a central role in the reproductive events associated with the establishment and maintenance of pregnancy. Progesterone receptor, a member of the steroid receptor superfamily, mediates the physiologic effects of progesterone. The PGR gene uses separate promoters and translational start sites to produce 2 isoforms, PRA and PRB, which are identical except for an additional 165 amino acids present only in the N terminus of PRB. Although PRA and PRB share several structural domains, they are distinct transcription factors that mediate their own response genes and physiologic effects with little overlap. It is composed of three domains: a modulating N terminal domain, a DNA binding domain and a C terminal steroid binding domain.
Progesterone levels 1. men 30-60 pg/0.1ml 2. women pre ovulatory phase: 20-160 pg/0.1ml; ovulatory phase: 1,000-1,700 pg/0.1ml; post ovulatory phase: 1,000-1,700 pg/0.1ml; Pregnant: 16-18 weeks: 300-800 pg/0.1ml; 28-30 weeks: 6,500-14,700 pg/0.1ml; 38-40 weeks: 12,000-19,000 pg/0.1ml.
Pregn 4 ene 3 20 dione antibody
has not yet been referenced specifically in any publications.