Western blot analysis was performed on whole cell extracts (30 ug lysate) of Hep G2 (Lane 1) and Mouse Pancreas (Lane 2). The blots were probed with Anti-Cytochrome P450 2A6 Rabbit polyclonal Antibody (Product# PA1342, 1:1000-1:5000 dilution dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G21234,1:5000 dilution). A 56 kDa band corresponding to Cytochrome P450 2A6 was observed in Hep G2 but not in Mouse Pancreas. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified, His-tagged, full length human P450 2A6 fusion protein.|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunohistochemistry (IHC)||See 1 publications below|
PA1-342 detects cytochrome P450 (P450) 2A6 from human, mouse and rat samples. This antibody detects, to a lesser extent, other P450 isoforms.
PA1-342 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~49 kDa protein representing P450 2A6 from rat liver extract.
The PA1-342 immunogen is purified, His-tagged, full length human P450 2A6 fusion protein.
The cytochrome P450 (P450) superfamily of enzymes is one of three enzyme systems which metabolize the fatty acid arachadonic acid (AA) to regulators of vascular tone. P450 enzymes are monooxygenase enzymes which require several co-factors such as nicotinamide adenine dinucleotide phosphate (NADPH) and P450 reductase. There are over 200 known genes which encode P450s. Epoxygenases are those P450s which metabolize AA to epoxyeicosatrienoic acid (EETs) and omega-hydroxylases are those P450s which produce 19- and 20-hydroxyeicosatetraenoic acids (19- and 20-HETE). As well as fatty acid metabolism, P450s also metabolize many drugs and toxins. Cytochrome P450 3A4 is abundantly expressed in liver and small intestine and is inducible by barbiturates, glucocorticoids and rifampicin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Expression and activity of cytochromes P450 2E1, 2A, and 2B in the mouse ovary: the effect of 4-vinylcyclohexene and its diepoxide metabolite.
PA1-342 was used in immunohistochemistry to characterize CYP450 isoforms in mouse ovary and to evaluate the effects of in vivo dosing with VCH and VCD on them
|Cannady EA,Dyer CA,Christian PJ,Sipes IG,Hoyer PB||Toxicological sciences : an official journal of the Society of Toxicology (73:423)||2003|