CYP11A1 Antibody

CYP11A1 Antibody is a Rabbit-derived and non-conjugated IgG polyclonal antibody, targeting to CYP11A1.

CYP11A1 is a cytochrome P450 monooxygenase that catalyzes the side-chain hydroxylation and cleavage of cholesterol to form pregnenolone, the precursor of most steroid hormones. The enzyme performs three sequential oxidation reactions on cholesterol: initial hydroxylation at C22, followed by hydroxylation at C20 to produce 20R,22R-hydroxycholesterol, which is subsequently cleaved between C20 and C22 to yield the C21-steroid pregnenolone and 4-methylpentanal. Mechanistically, it utilizes molecular oxygen, incorporating one oxygen atom into the substrate while reducing the second to water. The required two electrons are supplied by NADPH through a two-protein mitochondrial transfer system consisting of flavoprotein FDXR (adrenodoxin/ferredoxin reductase) and nonheme iron-sulfur protein FDX1 or FDX2 (adrenodoxin/ferredoxin).

P05108

Predicted band size: 60 kDa; Observed band size: 45 kDa

affinity purified

Mitochondrion membrane.

Non-conjugated

Unmodified

AB_3103283

Primary Antibody; Rabbit Polyclonal Antibody

Polyclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:1000; IHC: 1:50-1:100; IF: 1:50-1:200;

• 
  Western blot analysis of extracts from HepG2(lane 2(20ug) , NIH/3T3(lane 3(20ug) and HEK293(lane 4(20ug) using CYP11A1 Antibody (HY-P81223) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of HepG2 cells labeling CYP11A1 with CYP11A1 Antibody (HY-P81223)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with CYP11A1 Antibody (HY-P81223) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of HepG2 cells labeling CYP11A1 with CYP11A1 Antibody (HY-P81223) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with CYP11A1 Antibody (HY-P81223) at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded mouse liver tissue using CYP11A1 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded mouse liver tissue using CYP11A1 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-P, IHC-F, ICC/IF

Liquid

Supplied in 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

KLH conjugated synthetic peptide derived from human CYP11A1/P450SCC.