Cyclin T1 Antibody (monoclonal, 3B7)

• Catalog Number: orb763202
• Category: Antibodies
• Description: Cyclin T1 Antibody (monoclonal, 3B7)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 3B7
• Tested applications: FC, WB
• Reactivity: Human, Monkey
• Isotype: Mouse IgG2b
• Immunogen: A synthetic peptide corresponding to a sequence in the middle region of human Cyclin T1 (375-410aa QKQNSKSVPSAKVSLKEYRAKHAEELAAQKRQLENM), different from the related mouse sequence by one amino acid.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5 μg/ml, Human, Monkey Flow Cytometry, 1-3 μg/1x106 cells, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 81 kDa
• UniProt ID: O60563
• Storage: 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 freezing and thawing.
• 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. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Cyclin T1 using anti-Cyclin T1 antibody (orb763202). 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 Jurkat whole cell lysates, Lane 2: human HeLa whole cell lysates, Lane 3: human SW620 tissue lysates, Lane 4: monkey COS-7 whole cell 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-Cyclin T1 antigen affinity purified monoclonal antibody (Catalog # orb763202) 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 Cyclin T1 at approximately 81 kDa. The expected band size for Cyclin T1 is at 81 kDa.

Flow Cytometry analysis of U20S cells using anti-Cyclin T1 antibody (orb763202). Overlay histogram showing U20S cells stained with orb763202 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cyclin T1 Antibody (orb763202, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.