Immunofluorescence analysis of Cytochrome P450-3A4-3A5 was done on 70% confluent log phase HepG2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cytochrome P450 3A4 + 3A5 (F24 P2 B10) Mouse Monoclonal Antibody (MA3032) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Clone||F24 P2 B10|
|Immunogen||Synthetic peptide corresponding to residues E S R D G T V S G A, conjugated to ovalbumin.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||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.
|Immunohistochemistry (IHC)||See 1 publications below|
MA3-032 detects Cytochrome P450 3A4/3A5 from human cell line HepG2, and from bovine liver and pancreas samples.
MA3-032 has been successfully used in Western blot applications. By Western blot, this antibody detects a ~54 kDa band representing Cytochrome P450 3A4/3A5.
The MA3-032 immunogen is a synthetic peptide corresponding to residues E S R D G T V S G A, conjugated to ovalbumin.
This gene,CYP3A5, encodes a member of the cytochrome P450 superfamily of enzymes. The cytochrome P450 proteins are monooxygenases which catalyze many reactions involved in drug metabolism and synthesis of cholesterol, steroids and other lipids. This protein localizes to the endoplasmic reticulum and its expression is induced by glucocorticoids and some pharmacological agents. The enzyme metabolizes drugs such as nifedipine and cyclosporine as well as the steroid hormones testosterone, progesterone and androstenedione. This gene is part of a cluster of cytochrome P450 genes on chromosome 7q21.1. This cluster includes a pseudogene, CYP3A5P1, which is very similar to CYP3A5. This similarity has caused some difficulty in determining whether cloned sequences represent the gene or the pseudogene.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cytochrome p450 profile of colorectal cancer: identification of markers of prognosis.
MA3-032 was used in immunohistochemistry to investigate the changes of cytochrome p450 expression in colorectal tumors
|Kumarakulasingham M,Rooney PH,Dundas SR,Telfer C,Melvin WT,Curran S,Murray GI||Clinical cancer research : an official journal of the American Association for Cancer Research (11:3758)||2005|