rev: January 23, 2002
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& Related Antibodies
(anti-Human and others as indicated)
RDI Division of Fitzgerald Industries Intl offers a wide line of antibodies. Since no one antibody works best for all applications (neutralization, blotting, histochemistry, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.
Rabbit ANTI-HUMAN CYP3A4 IgG
Polyclonal Antibody Developed in Rabbits
Antiserum was developed in rabbits using purified human liver CYP3A4 as immunogen. The whole IgG fraction was purified from antiserum using caprylic acid/ammonium sulfate fractionation. Anti-human CYP3A4 IgG is provided as a powder after lyophylization from 100 mM potassium phosphate buffer (pH 7.4), 250 mM KCl, and 2.5 µM thimerosal (added as a preservative).
· Specificity and Purity
Specificity has been determining by Western blotting. Anti-human CYP3A4 IgG reacts primarily with CYP3A4 (51.5 kDa) but also recognizes CYP3A5 (52.5 kDa) and CYP3A7 (51.5 kDa) in human liver microsomes. This anti-CYP3A4 preparation inhibits reactions catalyzed by CYP3A proteins (e.g., testosterone 6b-hydroxylation and nifedipine oxidation). Cross-reaction of the antibody with homologous CYP3A proteins in liver microsomes from animal species has not been determined. Purity of the antibody has been established by SDS-PAGE run under denaturing conditions. Two protein bands of 50 kDa and 25 kDa are visualized by Coomassie blue staining, which represent the heavy and light chains, respectively, of rabbit IgG.
· Reconstitution and Storage
Reconstitute by adding 0.5 ml of distilled water to one vial of lyophilized IgG (1 mg). Rotate vial gently until powder dissolves. Add 0.5 ml of neat glycerol to obtain a 50% glycerol solution containing 1 mg IgG/ml. Prior to reconstitution, store the product at 0-5°C. After reconstitution, the solution can be stored at -20°C since the presence of 50% glycerol will prevent freeze/thaw cycles.
Immunoreactivity of anti-CYP3A4 IgG with human liver proteins
Lane A = UC9410 microsomes (10 µg)
Lane B = UC9001 microsomes (10 µg)
Lane C = UC9007 microsomes (10 µg)
Lane D = UC9402 microsomes (0.1 µg)
· Use for Western Blotting
1. For full size (15 x 15 cm) blots, add 200 µg rabbit anti-human CYP3A4 IgG to 15-20 ml of blocking solution (e.g., Blotto). This corresponds to 100 µl of the 1.0 mg IgG/ml solution prepared in 50% glycerol as described above. The amount can be scaled down proportionally with smaller size blots.
2. Incubate overnight at room temperature, and remove antibody solution (which can be stored at -20°C for re-use). After washing blot extensively with blocking solution to remove unbound CYP3A4 antibody, incubate with an anti-rabbit IgG conjugate of choice (e.g, anti-rabbit IgG peroxidase, anti-rabbit IgG-alkaline phosphatase or anti-rabbit IgG-biotin), and stain accordingly.
Note:we recommend titering for each use (ie, use of enhanced chemiluminescence detection systems may allow greater dilution of the antibody).
For In Vitro Research Use Only
Precaution: For in vitro research use only-not for use in or on humans or animals-not for use in diagnostics. Not responsible for any patent infringements with the use or derivation of these products.
See also antibodies against: CYP2A6, CYP2C8 CYP2C9, CYP2E1,CYP3A4 , CYP2C19 and CYP450 reductase.
Inquire for:pure native P450 enzymes
recombinant CYP450's now available!
RDI Division of Fitzgerald Industries Intl
34 Junction Square Drive
Concord MA 01742-3049
phone (978) 371-6446 or (800) 370-2222
fax (978) 371-2266
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