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Anti-CYP27A1 Rabbit Monoclonal Antibody

Figure 1. Western blot analysis of CYP27A1 using anti-CYP27A1 antibody (M02121-1).
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 HepG2 whole cell lysates,
Lane 2: human CACO-2 whole cell lysates,
Lane 3: rat liver tissue lysates,
Lane 4: mouse liver tissue 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-CYP27A1 antigen affinity purified monoclonal antibody (Catalog # M02121-1) at 1:500 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 1:500 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 CYP27A1 at approximately 50, 60 kDa. The expected band size for CYP27A1 is at 60 kDa.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC, FC
Primary Accession	Q02318
Host	Rabbit
Isotype	IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-CYP27A1 Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

Additional Information
Gene ID	1593
Other Names	Sterol 26-hydroxylase, mitochondrial, 1.14.15.15, 5-beta-cholestane-3-alpha, 7-alpha, 12-alpha-triol 26-hydroxylase, Cytochrome P-450C27/25, Cytochrome P450 27, Sterol 27-hydroxylase, Vitamin D(3) 25-hydroxylase, CYP27A1 {ECO:0000303\|PubMed:21411718, ECO:0000312\|HGNC:HGNC:2605}
Calculated MW	60235 Da
Application Details	WB 1:500-1:2000 IHC 1:50-1:200 ICC/IF 1:50-1:200 FC 1:80
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: 20C32
Immunogen	A synthesized peptide derived from human CYP27A1
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	CYP27A1 {ECO:0000303\|PubMed:21411718, ECO:0000312\|HGNC:HGNC:2605}
Function	Cytochrome P450 monooxygenase that catalyzes regio- and stereospecific hydroxylation of cholesterol and its derivatives. Hydroxylates (with R stereochemistry) the terminal methyl group of cholesterol side-chain in a three step reaction to yield at first a C26 alcohol, then a C26 aldehyde and finally a C26 acid (PubMed:12077124, PubMed:21411718, PubMed:28190002, PubMed:9660774). Regulates cholesterol homeostasis by catalyzing the conversion of excess cholesterol to bile acids via both the 'neutral' (classic) and the 'acid' (alternative) pathways (PubMed:11412116, PubMed:1708392, PubMed:2019602, PubMed:7915755, PubMed:9186905, PubMed:9660774, PubMed:9790667). May also regulate cholesterol homeostasis via generation of active oxysterols, which act as ligands for NR1H2 and NR1H3 nuclear receptors, modulating the transcription of genes involved in lipid metabolism (PubMed:12077124, PubMed:9660774). Catalyzes the conversion of 7-ketocholesterol (7-oxocholesterol) into 7-keto,27- hydroxycholesterol, a precursor of 7-keto,27-hydroxycholesterol, an active oxysterol that activates smoothened (SMO) and the smoothened signaling pathway (By similarity). Plays a role in cholestanol metabolism in the cerebellum. Similarly to cholesterol, hydroxylates cholestanol and may facilitate sterol diffusion through the blood-brain barrier to the systemic circulation for further degradation (PubMed:28190002). Also hydroxylates retinal 7-ketocholesterol, a noxious oxysterol with pro-inflammatory and pro-apoptotic effects, and may play a role in its elimination from the retinal pigment epithelium (PubMed:21411718). May play a redundant role in vitamin D biosynthesis. Catalyzes 25-hydroxylation of vitamin D3 that is required for its conversion to a functionally active form (PubMed:15465040).
Cellular Location	Mitochondrion inner membrane {ECO:0000250\|UniProtKB:P17178}; Peripheral membrane protein {ECO:0000250\|UniProtKB:P17178}. Note=Post-translationally targeted to mitochondria. All three of the receptor proteins in the TOM complex, TOMM70, TOMM20 and TOMM22 are required for the translocation across the mitochondrial outer membrane. After translocation into the matrix, associates with the inner membrane as a membrane extrinsic protein {ECO:0000250\|UniProtKB:P17178}
Tissue Location	Expressed in the neural retina and underlying retinal pigment epithelium (at protein level) (PubMed:21411718) Expressed in the gray and white matter of cerebellum (at protein level) (PubMed:28190002).