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CYP27A1 Recombinant Rabbit Monoclonal Antibody [JG40-36]

Usd: 385

Specification

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Catalog# ET7109-05

CYP27A1 Recombinant Rabbit Monoclonal Antibody [JG40-36]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

CYP27A1 Recombinant Rabbit Monoclonal Antibody [JG40-36]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human CYP27A1 aa 400-531 / 531.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 60 kDa

Positive Control

HepG2 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, HepG2, human liver cancer tissue, mouse liver tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

JG40-36

RRID

AB_3070895

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:100-1:1,000

Target

Function

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.

Background References

1. Cali J.J et al. Mutations in the bile acid biosynthetic enzyme sterol 27-hydroxylase underlie cerebrotendinous xanthomatosis. J Biol Chem 266:7779-7783 (1991).

2. Kim K-K et al. Identification of new mutations in sterol 27-hydroxylase gene in Japanese patients with cerebrotendinous xanthomatosis (CTX). J Lipid Res 35:1031-1039 (1994).

Sequence Similarity

Belongs to the cytochrome P450 family.

Tissue Specificity

Expressed in the neural retina and underlying retinal pigment epithelium (at protein level). Expressed in the gray and white matter of cerebellum (at protein level).

Subcellular Location

Mitochondrion membrane.

UNIPROT

SwissProt: Q02318 Human

SwissProt: Q9DBG1 Mouse

SwissProt: P17178 Rat

Synonyms

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

Expand

Images

  • Western blot analysis of CYP27A1 on different lysates with Rabbit anti-CYP27A1 antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/1,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (10 µg/Lane)<br />Lane 2: Mouse liver tissue lysate (20 µg/Lane)<br />Lane 3: Rat liver tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 60 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: Lane 1: 1 minute 2 seconds; Lane 2-3: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CYP27A1 on different lysates with Rabbit anti-CYP27A1 antibody (ET7109-05) 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)

    Predicted band size: 60 kDa
    Observed band size: 50 kDa

    Exposure time: Lane 1: 1 minute 2 seconds; Lane 2-3: 10 seconds; ECL: K1801;

    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 (ET7109-05) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling CYP27A1 with Rabbit anti-CYP27A1 antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-CYP27A1 antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/100 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-CYP27A1 antibody (ET7109-05) 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 (ET7109-05) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-CYP27A1 antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/100 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-CYP27A1 antibody (ET7109-05) 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 (ET7109-05) 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.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-CYP27A1 antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) at 1/100 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>) 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.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-CYP27A1 antibody (ET7109-05) 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 (ET7109-05) 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.

  • Flow cytometric analysis of HepG2 cells labeling CYP27A1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7109-05" style="font-weight: bold;text-decoration: underline;">ET7109-05</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling CYP27A1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET7109-05, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation