WB
Western blot analysis of CYP27A1 on different lysates with Rabbit anti-CYP27A1 antibody at 1/1,000 dilution. Lane 1: HepG2 cell lysate (10 µg/Lane), Lane 2: Mouse liver tissue lysate (20 µg/Lane), Lane 3: Rat liver tissue lysate (20 µg/Lane), Exposure time: Lane 1: 1 minute 2 seconds; Lane 2-3: 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-CYP27A1 antibody at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution for 1 hour at room temperature. 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-CYP27A1 antibody at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution for 1 hour at room temperature. 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.IHC
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-CYP27A1 antibody at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution for 1 hour at room temperature. 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.ICC/IF
Immunocytochemistry analysis of HepG2 cells labeling CYP27A1 with Rabbit anti-CYP27A1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CYP27A1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.FC
Flow cytometric analysis of HepG2 cells labeling CYP27A1. Cells were fixed and permeabilized. Then stained with the primary antibody (1/100) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | CYP27A1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | P450 enzymes constitute a family of monooxygenase enzymes that are involved in the metabolism of a wide array of endo-genous and xenobiotic compounds. P450 enzymes can be classified, based on their sequence similarities, into distinct subfamilies, which include CYP1A and CYP2A. Other P450 family members include CYP19, also designated aromatase (P450arom), which catalyzes the conversion of C19 steroids to estrogens in various tissues, including placenta, gonads, adipose tissue, skin and brain. CYP19 expression is controlled by hormonally regulated promoters in different tissues and increased aromatase activity is associated with familial gynecomastia. Also, a polymorphic allele of CYP19 (repeat (TTTA)12) is present in a majority of breast cancer patients. P450 cholesterol 7a-hydroxylase, CYP7A1, is the rate limiting enzyme of bile acid synthesis in the liver, and its expression is mediated by the bile acid receptor FXR. CYP27A1 catalyzes vitamin D3 25-hydroxylation and is localized to the mitochondria in kidney and liver. |
| Immunogen | Recombinant protein within Human CYP27A1 aa 400-531 / 531. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:500-1:2000 IHC:1:50-1:200 ICC/IF:1:50-1:200 FC:1:100 |
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| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | 12-alpha-triol 27-hydroxylase antibody
5-beta-cholestane-3-alpha antibody
5-beta-cholestane-3-alpha 7-alpha 12-alpha-triol 26-hydroxylase antibody 5-beta-cholestane-3-alpha 7-alpha 12-alpha-triol 27-hydroxylase antibody 5-beta-cholestane-3-alpha 7-alpha 12-alpha-triol 27-hydroxylase antibody 7-alpha antibody Cholestanetriol 26 monooxygenase antibody CP27 antibody CP27A_HUMAN antibody CTX antibody CYP antibody CYP27 antibody CYP27A1 antibody Cytochrome P 450C27/25 antibody Cytochrome P-450C27/25 antibody Cytochrome P450 27 antibody Cytochrome P450 27 mitochondrial antibody Cytochrome P450 family 27 subfamily A member 1 antibody Cytochrome P450 family 27 subfamily A polypeptide 1 antibody Cytochrome P450 subfamily XXVIIA (steroid 27-hydroxylase cerebrotendinous xanthomatosis) polypeptide 1 antibody mitochondrial antibody Sterol 26 hydroxylase antibody Sterol 26 hydroxylase mitochondrial antibody Sterol 26-hydroxylase antibody Sterol 27 hydroxylase antibody Sterol 27-hydroxylase antibody Vitamin D(3) 25 hydroxylase antibody Vitamin D(3) 25-hydroxylase antibody |
|---|---|
| Molecular Weight(MW) | 60kDa(Observed band size: 50kDa) |
| Cellular Localization | Mitochondrion membrane. |
WB
Western blot analysis of CYP27A1 on different lysates with Rabbit anti-CYP27A1 antibody at 1/1,000 dilution. Lane 1: HepG2 cell lysate (10 µg/Lane), Lane 2: Mouse liver tissue lysate (20 µg/Lane), Lane 3: Rat liver tissue lysate (20 µg/Lane), Exposure time: Lane 1: 1 minute 2 seconds; Lane 2-3: 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-CYP27A1 antibody at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution for 1 hour at room temperature. 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-CYP27A1 antibody at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution for 1 hour at room temperature. 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-CYP27A1 antibody at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution for 1 hour at room temperature. 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.
ICC/IF
Immunocytochemistry analysis of HepG2 cells labeling CYP27A1 with Rabbit anti-CYP27A1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CYP27A1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
FC
Flow cytometric analysis of HepG2 cells labeling CYP27A1. Cells were fixed and permeabilized. Then stained with the primary antibody (1/100) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:500-1:2000 IHC:1:50-1:200 ICC/IF:1:50-1:200 FC:1:100 |
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| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
