Anti-HSD3B1 Antibody (monoclonal, 3F3C/7F9C8)

• Catalog Number: orb865663
• Category: Antibodies
• Description: Anti-HSD3B1 Antibody (monoclonal, 3F3C/7F9C8). Tested in IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 3F3C/7F9C8
• Tested applications: ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Mouse IgG2b
• Immunogen: E.coli-derived human HSD3B1 recombinant protein (Position: Q17-Q373).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 42 kDa
• UniProt ID: P14060
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: chromosomal protein, Nonhistone, HMG4 antibody; Hi
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of HSD3B1 using anti-HSD3B1 antibody. HSD3B1 was detected in an immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-HSD3B1 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse 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 HSD3B1 using anti-HSD3B1 antibody. 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 placenta tissue lysates, Lane 2: rat testis tissue lysates, Lane 3: mouse testis 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 mouse anti-HSD3B1 antigen affinity purified monoclonal antibody 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for HSD3B1 at approximately 42 kDa. The expected band size for HSD3B1 is at 42 kDa.