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Western Blot (WB) (Figure 1. Western blot analysis of CYP7A1 using anti-CYP7A1 antibody (MBS1753447).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 50ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: mouse liver tissue lysatesLane 3: monkey liver tissue lysatesLane 4: human HepG2 whole cell lysatesLane 5: human Hele whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 # MBS1753447) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for CYP7A1 at approximately 54KD. The expected band size for CYP7A1 is at 54KD. )

Rabbit CYP7A1 Polyclonal Antibody | anti-CYP7A1 antibody

Anti-CYP7A1 Antibody

Gene Names
CYP7A1; CP7A; CYP7; CYPVII
Reactivity
Human, Mouse, Rat, Monkey
Applications
Western Blot, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
CYP7A1; Polyclonal Antibody; Anti-CYP7A1 Antibody; CYP7; Cytochrome P450 7A1; 24-hydroxycholesterol 7-alpha-hydroxylase; EC 1. 14. 14. 26; CYPVII; Cholesterol 7-alpha-hydroxylase; Cholesterol 7-alpha-monooxygenase; EC 1. 14. 14. 23; cytochrome P450 family 7 subfamily A member 1; anti-CYP7A1 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat, Monkey
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for CYP7A1 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Applicable Applications for anti-CYP7A1 antibody
Western Blot (WB), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml|Human, Mouse, Rat, Monkey|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Immunogen
E Coli-derived human CYP7A1 recombinant protein (Position: I10-L504).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of CYP7A1 using anti-CYP7A1 antibody (MBS1753447).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 50ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: mouse liver tissue lysatesLane 3: monkey liver tissue lysatesLane 4: human HepG2 whole cell lysatesLane 5: human Hele whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 # MBS1753447) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for CYP7A1 at approximately 54KD. The expected band size for CYP7A1 is at 54KD. )

Western Blot (WB) (Figure 1. Western blot analysis of CYP7A1 using anti-CYP7A1 antibody (MBS1753447).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 50ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: mouse liver tissue lysatesLane 3: monkey liver tissue lysatesLane 4: human HepG2 whole cell lysatesLane 5: human Hele whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 # MBS1753447) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for CYP7A1 at approximately 54KD. The expected band size for CYP7A1 is at 54KD. )

Flow Cytometry (FC/FACS)

(Figure 2. Flow Cytometry analysis of U87 cells using anti-CYP7A1 antibody (MBS1753447).Overlay histogram showing U87 cells stained with MBS1753447 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CYP7A1 Antibody (MBS1753447, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 2. Flow Cytometry analysis of U87 cells using anti-CYP7A1 antibody (MBS1753447).Overlay histogram showing U87 cells stained with MBS1753447 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CYP7A1 Antibody (MBS1753447, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-CYP7A1 antibody
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.
References
1. Agellon, L. B., Drover, V. A. B., Cheema, S. K., Gbaguidi, G. F., Walsh, A. Dietary cholesterol fails to stimulate the human cholesterol 7-alpha-hydroxylase gene (CYP7A1) in transgenic mice. J. Biol. Chem. 277: 20131-20134, 2002.
2. Cohen, J. C., Cali, J. J., Jelinek, D. F., Mehrabian, M., Sparkes, R. S., Lusis, A. J., Russell, D. W., Hobbs, H. H. Cloning of the human cholesterol 7-alpha-hydroxylase gene (CYP7) and localization to chromosome 8q11-q12. Genomics 14: 153-161, 1992.
3. Molowa, D. T., Chen, W. S., Cimis, G. M., Tan, C. P. Transcriptional regulation of the human cholesterol 7 alpha-hydroxylase gene. Biochemistry 31: 2539-2544, 1992.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
57,661 Da
NCBI Official Full Name
cholesterol 7-alpha-monooxygenase
NCBI Official Synonym Full Names
cytochrome P450, family 7, subfamily A, polypeptide 1
NCBI Official Symbol
CYP7A1
NCBI Official Synonym Symbols
CP7A; CYP7; CYPVII
NCBI Protein Information
cholesterol 7-alpha-monooxygenase; cytochrome P450 7A1; cholesterol 7-alpha-hydroxylase; cytochrome P450, subfamily VIIA polypeptide 1
UniProt Protein Name
Cholesterol 7-alpha-monooxygenase
UniProt Gene Name
CYP7A1
UniProt Synonym Gene Names
CYP7
UniProt Entry Name
CP7A1_HUMAN

NCBI Description

This gene encodes a member of the cytochrome P450 superfamily of enzymes. The cytochrome P450 proteins are monooxygenases which catalyze many reactions involved in drug metabolism and synthesis of cholesterol, steroids and other lipids. This endoplasmic reticulum membrane protein catalyzes the first reaction in the cholesterol catabolic pathway in the liver, which converts cholesterol to bile acids. This reaction is the rate limiting step and the major site of regulation of bile acid synthesis, which is the primary mechanism for the removal of cholesterol from the body. Polymorphisms in the promoter of this gene are associated with defects in bile acid synthesis. [provided by RefSeq, Feb 2010]

Uniprot Description

CYP7A1: Catalyzes a rate-limiting step in cholesterol catabolism and bile acid biosynthesis by introducing a hydrophilic moiety at position 7 of cholesterol. Important for cholesterol homeostasis. Belongs to the cytochrome P450 family.

Protein type: Lipid Metabolism - primary bile acid biosynthesis; Endoplasmic reticulum; Oxidoreductase; EC 1.14.13.17

Chromosomal Location of Human Ortholog: 8q11-q12

Cellular Component: endoplasmic reticulum membrane; intracellular membrane-bound organelle

Molecular Function: cholesterol 7-alpha-monooxygenase activity; iron ion binding; heme binding

Biological Process: bile acid biosynthetic process; cholesterol homeostasis; bile acid metabolic process; xenobiotic metabolic process; cellular lipid metabolic process; cholesterol catabolic process; sterol metabolic process; transmembrane transport

Research Articles on CYP7A1

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Product Notes

The CYP7A1 cyp7a1 (Catalog #AAA1753447) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-CYP7A1 Antibody reacts with Human, Mouse, Rat, Monkey and may cross-react with other species as described in the data sheet. AAA Biotech's CYP7A1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human, Mouse, Rat, Monkey| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the CYP7A1 cyp7a1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CYP7A1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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