Western blot analysis was performed on whole cell extracts (30 ug lysate) of Mouse Liver (Lane 1) and Rat Liver (Lane 2). The blots were probed with Anti-Cytochrome P450 3A4 Rabbit Polyclonal Antibody (Product# PA1343, 1:1000-1:5000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G21234,1:5000 dilution). A 57 kDa band corresponding to Cytochrome P450 3A4 was observed across tissues tested. 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, Rat|
|Published species reactivity||Rat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified, His-tagged, full length human P450 3A4 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.
PA1-343 detects cytochrome P450 (P450) 3A4 from human and rat tissues and cells. This antibody detects, to a lesser extent, other P450 isoforms.
PA1-343 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~46 kDa protein representing P450 3A4 from rat liver extracts.
The PA1-343 immunogen is purified, His-tagged, full length human P450 3A4 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.
Effects of lersivirine on canine and rodent thyroid function.
PA1-343 was used in immunohistochemistry to study the effects of the non-nucleotide reverse transcriptase inhibitor lersivirine on thyroid function in animals
|Houle CD,Finch GL,Mauthe RJ,Potter DM,Walisser JA,Gardner IB,DeWit RH||Toxicologic pathology (42:897)||2014|
Drug-metabolizing enzyme and transporter expression in a mouse model of diabetes and obesity.
PA1-343 was used in western blot to investigate the expression of drug-metabolizing enzymes and transporters in diabetic mouse model
|Cheng Q,Aleksunes LM,Manautou JE,Cherrington NJ,Scheffer GL,Yamasaki H,Slitt AL||Molecular pharmaceutics (5:77)||2008|