Product nameAnti-Cytochrome P450 1A1 antibody
See all Cytochrome P450 1A1 primary antibodies
DescriptionRabbit polyclonal to Cytochrome P450 1A1
SpecificityDetects Cytochrome P450 1A1 from Human cells. This antibody detects, to a lesser extent, other P450 isoforms.
Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
Species reactivityReacts with: Human
Other Immunogen Type (His-tag) corresponding to Human Cytochrome P450 1A1. Purified, His-tagged, full length human P450 1A1 fusion protein.
- MCF-7 cell lysate.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituent: 99% PBS
Concentration information loading...
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
Our Abpromise guarantee covers the use of ab3568 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
By Western blot, this antibody detects an ~54 kDa protein representing P450 1A1 from MCF-7 cell extract.
FunctionCytochromes P450 are a group of heme-thiolate monooxygenases. In liver microsomes, this enzyme is involved in an NADPH-dependent electron transport pathway. It oxidizes a variety of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics.
Tissue specificityLung, lymphocytes and placenta.
Sequence similaritiesBelongs to the cytochrome P450 family.
Cellular localizationEndoplasmic reticulum membrane. Microsome membrane.
- Information by UniProt
- AHH antibody
- AHRR antibody
- Aryl hydrocarbon hydroxylase antibody
All lanes : Anti-Cytochrome P450 1A1 antibody (ab3568) at 1/500 dilution
Lane 1 : HepG2 with Skimmed milk
Lane 2 : LNCaP with Skimmed milk
Lane 3 : K562 with Skimmed milk
Lane 4 : CaCo-2 with Skimmed milk
Lane 5 : HeLa with Skimmed milk
Lysates/proteins at 30 µg per lane.
Blocking peptides at 5 % per lane.
All lanes : Goat anti-rabbit (H+L), HRP conjugated at 1/5000 dilution
ab3568 staining Cytochrom P450 1A1 in HepG2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/250 in 0.1% BSA) for 3 hours at room temperature (panel a). An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG (H+L) polyclonal was used as the secondary antibody (1/2000). Nuclei were stained blue with DAPI (panel b), F-actin stained with Rhodamine Phalloidin (panel c) and merged image showing cyctoplasmic localization (panel d)
HepG2 cells were incubated at 37°C for 24h with vehicle control (0 µM) and different concentrations of itraconazole (ab120816). Increased expression of cytochrome P450 1A1 (ab3568) in HepG2 cells correlates with an increase in nifuroxazide concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab3568 at 1/500 dilution and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
IHC image of ab3568 staining in Human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3568, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab3568 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3568, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab3568 at a dilution of 1/500 staining MCF-7 cell extract by Western blot.
HepG2 cells were incubated at 37°C for 24h with vehicle control (0 µM) and 100 µM of fluconazole (ab141065). Increased expression of cytochrome P450 1A1 (ab3568) correlates with an increase in fluconazole concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab3568 at 1/500 dilution and ab8227 at 1 µg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
This product has been referenced in:
- Alarcan J et al. Metabolism of the Marine Phycotoxin PTX-2 and Its Effects on Hepatic Xenobiotic Metabolism: Activation of Nuclear Receptors and Modulation of the Phase I Cytochrome P450. Toxins (Basel) 9:N/A (2017). WB ; Human . Read more (PubMed: 28678150) »
- Mitsui Y et al. Functional role and tobacco smoking effects on methylation of CYP1A1 gene in prostate cancer. Oncotarget 7:49107-49121 (2016). Read more (PubMed: 27203547) »