|Product Name||Anti-Cytochrome P450 2E1/CYP2E1 Picoband™ Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Cytochrome P450 2E1(CYP2E1) detection. Tested with WB, IHC-P in Human;Mouse;Rat.|
|Cite This Product||Anti-Cytochrome P450 2E1/CYP2E1 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9190)|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.|
|Immunogen||E.coli-derived human CYP2E1 recombinant protein (Position: M1-Y310). Human CYP2E1 shares 73% and 74% amino acid (aa) sequences identity with mouse and rat CYP2E1, respectively.|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat, Human, By Heat
Western blot, 0.1-0.5μg/ml, Mouse, Rat, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure 1. Western blot analysis of CYP2E1 using anti-CYP2E1 antibody (PB9190).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
lane 1: rat liver tissue lysate,
lane 2: mouse liver tissue lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CYP2E1 antigen affinity purified polyclonal antibody (Catalog # PB9190) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CYP2E1 at approximately 55KD. The expected band size for CYP2E1 is at 57KD.
Figure 2. IHC analysis of CYP2E1 using anti-CYP2E1 antibody (PB9190).
CYP2E1 was detected in paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CYP2E1 Antibody (PB9190) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Figure 3. IHC analysis of CYP2E1 using anti-CYP2E1 antibody (PB9190).
CYP2E1 was detected in paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CYP2E1 Antibody (PB9190) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Cytochrome P450 2E1|
|Alternative Names||Cytochrome P450 2E1;1.14.13.-;4-nitrophenol 2-hydroxylase;1.14.13.n7;CYPIIE1;Cytochrome P450-J;Cytochrome P450 2E1, N-terminally processed;CYP2E1;CYP2E;|
|Subcellular Localization||Endoplasmic reticulum membrane; Peripheral membrane protein. Microsome membrane; Peripheral membrane protein.|
|Molecular Weight||56849 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Metabolizes several precarcinogens, drugs, and solvents to reactive metabolites. Inactivates a number of drugs and xenobiotics and also bioactivates many xenobiotic substrates to their hepatotoxic or carcinogenic forms.|
|Research Areas||Cancer, Cardiovascular, Cytochromes, Drug Metabolism, Lipases, Lipid And Lipoprotein Metabolism, Lipid Metabolism, Lipids / Lipoproteins, Metabolic Signaling Pathways, Metabolism, Mitochondrial Metabolism, Pathways And Processes, Signal Transduction
*You can search these to find other products in these research areas.
|Background||Cytochrome P450 2E1 (abbreviated CYP2E1), a member of the cytochrome P450 mixed-function oxidase system, is involved in the metabolism of xenobiotics in the body. In humans, the CYP2E1 enzyme is encoded by the CYP2E1 gene. It is mapped to 10q26.3. While it is involved in the oxidative metabolism of a small range of substrates (mostly small polar molecules), there are many important drug interactions mediated by CYP2E1. Most drugs undergo deactivation by CYP2E1, either directly or by facilitated excretion from the body. Also, many substances are bioactivated by CYP2E1 to form their active compounds. In addition, CYP2E1 is an important enzyme for the conversion of ethanol to acetaldehyde and to acetate in humans. In the conversion sequence of acetyl-CoA to glucose, CYP2E1 transforms acetone via acetol into propylene glycol and methylglyoxal, the precursors of pyruvate, acetate and lactate.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,