CYP27A1 Recombinant Rabbit Monoclonal Antibody [JG40-36]
cat.: ET7109-05
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC |
| Clonality: | Monoclonal |
| Clone number: | JG40-36 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | 60 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human CYP27A1 aa 400-531 / 531. |
| Positive control: | Rat heart tissue lysate, mouse liver tissue lysate, human liver tissue lysate, HepG2, human liver carcinoma tissue, human kidney tissue, human small intestine tissue, Hela. |
| Subcellular location: | Mitochondrion membrane. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:500-1:2,000 1:50-1:200 1:50-1:200 1:50-1:100 |
| Uniprot #: | SwissProt: Q02318 Human | Q9DBG1 Mouse | P17178 Rat |
| Alternative names: | 12-alpha-triol 27-hydroxylase 5-beta-cholestane-3-alpha 5-beta-cholestane-3-alpha, 7-alpha, 12-alpha-triol 26-hydroxylase 5-beta-cholestane-3-alpha, 7-alpha, 12-alpha-triol 27-hydroxylase 5-beta-cholestane-3-alpha,7-alpha,12-alpha-triol 27-hydroxylase 7-alpha Cholestanetriol 26 monooxygenase CP27 CP27A_HUMAN CTX CYP CYP27 CYP27A1 Cytochrome P 450C27/25 Cytochrome P-450C27/25 Cytochrome P450 27 Cytochrome P450 27 mitochondrial Cytochrome P450 family 27 subfamily A member 1 Cytochrome P450 family 27 subfamily A polypeptide 1 Cytochrome P450 subfamily XXVIIA (steroid 27-hydroxylase cerebrotendinous xanthomatosis) polypeptide 1 mitochondrial Sterol 26 hydroxylase Sterol 26 hydroxylase mitochondrial Sterol 26-hydroxylase Sterol 27 hydroxylase Sterol 27-hydroxylase Vitamin D(3) 25 hydroxylase Vitamin D(3) 25-hydroxylase |
Images
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Fig1:
Western blot analysis of CYP27A1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7109-05, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Rat heart tissue lysate Lane 2: Mouse liver tissue lysate Lane 3: Human liver tissue lysate |
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Fig2:
Immunocytochemistry analysis of HepG2 cells labeling CYP27A1 with Rabbit anti-CYP27A1 antibody (ET7109-05) 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 (ET7109-05) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. |
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Fig3: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-CYP27A1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-05, 1/50) for 30 minutes 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. |
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Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-CYP27A1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-05, 1/50) for 30 minutes 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. |
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Fig5: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-CYP27A1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-05, 1/50) for 30 minutes 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. |
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Fig6: Flow cytometric analysis of CYP27A1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7109-05, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow). |
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