rev:  Janaury 23, 2002

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ANTI-CYTOCHROME P450

& Related Antibodies  

(anti-Human and others as indicated)

RDI Divison 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.


ANTI-CYP2C19


Rabbit anti-human Cytochrome P450 CYP2C19   NEW MONOSPECIFIC ANTIBODY



Rabbit anti-human Cytochrome P450 CYP2C19 NEW FORMULATION   MONOSPECIFIC


CAT# RDI-CYP2C19Mabr


 Price: $1000.00/1mg vial

       
Presentation: 1 milligram lyophilized from 100mM potassium phosphate buffer pH 7.4, 250mM KCL and 2.5 micromole thimerosal.


species: rabbit IgG (purified from serum using caprylic acid/ammonium sulfate fractionation.


Sample Blotting Data:

blotting picture of
anti-cyp2c19m



1= UC8905 Liver Mix   20ug          Note: Top Dark Bands on all lanes are non specific keratin staining

2=CYP2C19                 0.1ug         Unasborbed tested with 150ug/20ml of antibody, (7.5ug/ml)

3=CYP2C8                   0.1ug          -absorbed antibody at 300ug/20ml  (15ug/ml)

4=CYP2C9                   0.1ug          -titer for each detection sysem

5=UC9408 Liver Mix     20ug          -MW CYP2C19= 50 kDa,  CYP2C9=56 kDa, CYP2C8=51.5kDa

6=UC9411 Liver Mix     20ug


Specificity has been determining by Western blotting. Anti-human CYP2C19 IgG reacts only with its 50.5 kDa immunogen in human liver microsomes. Cross-reactivity with CYP2C8 and CYP2C9 has removed by back-adsorption against these microsomal proteins coupled to Sepharose beads. Reactivity of anti-CYP2C19 with CYP2C proteins in rat, rabbit, and hamster liver microsomes is unknown. Purity of the antibody has been established by SDS-PAGE run under denaturing conditions. Two protein bands with molecular weights of 50 kDa and 25 kDa can be visualized by Coomassie blue staining, which correspond to the heavy and light chains, respectively, of rabbit IgG.


Reconstitution: Immunoblots: Reconstitute by adding 0.5ml of   distilled water to one vial of lyophilized IgG (1mg). Rotate vial gently until powder dissolves. Add 0.5ml of neat glycerol to obtain a 50% glycerol solution containing 1mg IgG/ml. Store -20 DEG C.

Immunoinhibition: Reconstitute the antibodies in 100 microliters of the assay buffer being used so that the final concentration of antibody is 10mg/ml. Store at -20 DEG C. Avoid frequent freeze thaw cycles.


Note: All uses and titers must be optimized for each application.


Sample Protocol: western blot:for full size (15 X 15cm) blots, add 300ug rabbit anti-human CYP2C19M IgG to 15-20ml of blocking solution (eg., Blotto). This corresponds to 300ul of the 1.0mg IgG/ml solution prepared in 50% glycerol as described above. This amount can be scaled down appropriately with smaller size blots.


Incubate overnight at room temperature and then discard antibody solution. Wash blot extensively with blocking solution to remove unbound CYP2C19M antibody and then incubate with an anti-rabbit IgG conjugate (eg. anti-rabbit IgG-alkaline phosphatase or anti- rabbit IgG-biotin.)


In the developers lab, they use a goat anti- rabbit IgG biotin conjugate followed by a complex of Streptavidin-horse radish peroxidase and subsequently staining with 4- chloro-1-napththol/hydrogen peroxide. This system allows good visualization of CYP2C19 with 15-20ug of human liver microsomal protein applied to the original polyacrylamide gel However, an anti-rabbit IgG-alkaline phosphatase conjugate with chemiluminescent staining should be at least as sensitive.


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 (in production), CYP2C19M ,CYP450 reductase, CYP450 side chain cleavage.


Inquire for:pure natural P450 enzymes


RDI Divison of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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