CYP27A1 Antibody

• Catalog Number: orb654418
• Category: Antibodies
• Description: CYP27A1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human CYP27A1 recombinant protein (Position: A7-C531).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.25μg/ml, Human, Mouse, Monkey, Rat Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Direct ELISA, 0.1-0.5μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 60 kDa
• UniProt ID: Q02318
• Storage: 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.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

IF analysis of CYP27A1 using anti-CYP27A1 antibody (orb654418). CYP27A1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-CYP27A1 Antibody (orb654418) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of CYP27A1 using anti-CYP27A1 antibody (orb654418). 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: human placenta tissue lysates, Lane 2: human HEPG2 whole cell lysates, Lane 3: human CACO-2 whole cell lysates, Lane 4: human SW620 whole cell lysates, Lane 5: human SGC-7901 whole cell lysates, Lane 6: human HEK293 whole cell lysates, Lane 7: monkey liver tissue lysates, Lane 8: human U20S whole cell lysates, Lane 9: rat liver tissue lysates, Lane 10: rat heart tissue lysates, Lane 11: mouse liver tissue lysates, Lane 12: mouse heart tissue lysates, Lane 13: mouse NIH/3T3 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-CYP27A1 antigen affinity purified polyclonal antibody (Catalog # orb654418) at 0.25 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 # orb90503) with Tanon 5200 system. A specific band was detected for CYP27A1 at approximately 60KD. The expected band size for CYP27A1 is at 60KD.