Anti-CYP7A1 Antibody

Product Name

Anti-CYP7A1 Antibody

View all CYP7A1 Antibodies

SKU/Catalog Number

RP1079

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CYP7A1 Antibody catalog # RP1079. Tested in Flow Cytometry, WB applications. This antibody reacts with Human, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-CYP7A1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1079)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human CYP7A1 recombinant protein (Position: M301-L504). Human CYP7A1 shares 85.2 % and 83.3% amino acid (aa) sequence identity with mouse and rat CYP7A1, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

RP1079 is reactive to CYP7A1 in Human, Rat

Applications

RP1079 is guaranteed for Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

58 kDa

Calculated molecular weight

57661 MW

Background of CYP7A1

CYP7A1 (Cytochrome P450 Subfamily VIIA Polypeptide 1), also called CYP7 or CHOLESTEROL 7-ALPHA -HYDROXYLASE, is an enzyme that in humans is encoded by the CYP7A1 gene. Using both mouse-human somatic cell hybrids and in situ chromosomal hybridization, Cohen et al. (1992) mapped the CYP7 gene to 8q11-q12. By transfection of reporter constructs, mutation analysis, and DNase footprinting, Molowa et al. (1992) identified areas of the CYP7A1 promoter region that showed hepatocyte-specific activation. They found HNF3 to be an activator of CYP7A1 activity. Agellon et al. (2002) found that wildtype mice and mice transgenic for human CYP7A1 respond differently to cholesterol feeding. Cholesterol feeding stimulated Cyp7a1 mRNA abundance and enzymatic activity in wildtype mice, but repressed human CYP7A1 mRNA and activity in transgenic mice.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconstitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Rat
Flow Cytometry (Fixed), 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CYP7A1 using anti-CYP7A1 antibody (RP1079).
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 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: rat RH35 whole clel lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-CYP7A1 antigen affinity purified polyclonal antibody (Catalog # RP1079) 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 1:5000 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 CYP7A1 at approximately 58 kDa. The expected band size for CYP7A1 is at 58 kDa.

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Figure 2. Flow Cytometry analysis of SiHa cells using anti-CYP7A1 antibody (RP1079).
Overlay histogram showing SiHa cells stained with RP1079 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CYP7A1 Antibody (RP1079, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Gene/Protein Information For CYP7A1 (Source: Uniprot.org, NCBI)

Alternative Names

Cholesterol 7-alpha-hydroxylase; cholesterol 7-alpha-monooxygenase; CP7A; CYP7; CYP7A1; CYP7MGC126826; CYPVII; Cytochrome P450 7A1; cytochrome P450, family 7, subfamily A, polypeptide 1; cytochrome P450, subfamily VIIA (cholesterol 7 alpha-monooxygenase); cytochrome P450, subfamily VIIA polypeptide 1; EC 1.14.13; EC 1.14.13.17; MGC138389; polypeptide 1 CYP7A1 CP7A, CYP7, CYPVII cytochrome P450 family 7 subfamily A member 1 cytochrome P450 7A1|24-hydroxycholesterol 7-alpha-hydroxylase|cholesterol 7-alpha-hydroxylase|cholesterol 7-alpha-monooxygenase|cholesterol 7alpha-hydroxylase|cytochrome P450, family 7, subfamily A, polypeptide 1|cytochrome P450, subfamily VIIA polypeptide 1

For more info on CYP7A1, check out the CYP7A1 Infographic

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In this infographic, you will see the following information for CYP7A1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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