Immunofluorescent analysis of Cytochrome P450 2C11 (green) showing staining in the membrane of H-4-II-E cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cytochrome P450 2C11 polyclonal antibody (Product # PA3-034) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Cat, Human, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptides corresponding to residues D(49) I G Q S I K K F S K V(60) and Q(491) R A D S L S S H L(500) of rat Cytochrome P450 2C11.|
|Storage buffer||whole serum diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:500|
|Western Blot (WB)||1:1,500|
* 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.
|ELISA (ELISA)||See 2 publications below|
PA3-034 detects cytochrome P450 2C11 (P450 2C11) from human, feline and rat tissues.
PA3-034 has been successfully used in Western blot, ICC/IF and immunohistochemical (frozen and paraffin) procedures. By Western blot, this antibody detects an ~50 kDa protein representing P450 2C11 from rat liver extract. Immunohistochemical staining of P450 2C11 in rat brain with PA3-034 results in perivascular staining.
The PA3-034 immunogen is a synthetic peptides corresponding to residues D(49) I G Q S I K K F S K V(60) and Q(491) R A D S L S S H L(500) of rat Cytochrome P450 2C11.
The cytochrome P450 (P450) family 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 NADPH and P450 reductase. There are over 200 cDNA and quote;s which encode P450 protein. Epoxygenases are those P450 proteins which metabolize AA to epoxyeicosatrienoic acid (EETs) and omega-hydroxylases are those P450 proteins which produce 19- and 20-hydroxyeicosatetraenoic acids (19- and 20-HETE).
EET and quote;s, which exhibit vasodilation activity, are formed when an epoxide group is inserted between the unsaturated carbons of AA in positions 5,6; 8,9; 11,12; 14,15. EET and quote;s are produced in cerebral cortical tissue, coronary arteries and vascular endothelium. EET and quote;s are converted from AA by the 2C11 family of P450 and quote;s whose expression is induced by testosterone and is therefore not generally found in females.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Short-term inhibitory effects of nitric oxide on cytochrome P450-mediated drug metabolism: time dependency and reversibility profiles in isolated perfused rat livers.
PA3-034 was used in ELISA to study the dynamics and the reversibility of the effect of NO on P450s by means of the model of perfused rat livers
|Vuppugalla R,Mehvar R||Drug metabolism and disposition: the biological fate of chemicals (32:1446)||2004|
Hepatic disposition and effects of nitric oxide donors: rapid and concentration-dependent reduction in the cytochrome P450-mediated drug metabolism in isolated perfused rat livers.
PA3-034 was used in ELISA to study the impact of NO donors on the down-regulation of CYP450 without cytokines or other confounding in vivo factors
|Vuppugalla R,Mehvar R||The Journal of pharmacology and experimental therapeutics (310:718)||2004|