PROM1/CD133 Antibody (monoclonal, 8B6)

• Catalog Number: orb692235
• Category: Antibodies
• Description: PROM1/CD133 Antibody (monoclonal, 8B6)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 8B6
• Tested applications: FC, WB
• Reactivity: Human, Mouse
• Isotype: Mouse IgG2a
• Immunogen: E.coli-derived human PROM1/CD133 recombinant protein (Position: P531-H865). HumanPROM1/CD133 shares 61% amino acid (aa) sequence identity with mouse PROM1/CD133.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5μg/ml, Human, Mouse Flow Cytometry, 1-3μg/1x106 cells, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 120 kDa
• UniProt ID: O43490
• 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.

Western blot analysis of PROM1/CD133 using anti-PROM1/CD133 antibody (orb692235). 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 CACO-2 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: mouse kidney tissue 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 mouse anti-PROM1/CD133 antigen affinity purified monoclonal antibody (Catalog # orb692235) 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for PROM1/CD133 at approximately 120KD. The expected band size for PROM1/CD133 is at 120KD.

Flow Cytometry analysis of CACO-2 cells using anti-PROM1/CD133 antibody (orb692235). Overlay histogram showing CACO-2 cells stained with orb692235 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PROM1/CD133 Antibody (orb692235, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.