TRAP/CD40L/CD40LG Antibody

• Catalog Number: orb745937
• Category: Antibodies
• Description: TRAP/CD40L/CD40LG Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human TRAP/CD40L/CD40LG recombinant protein (Position: E76-H249).
• 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, Rat Flow Cytometry, 1-3μg/1x106 cells, Human, Rat Direct ELISA, 0.1-0.5μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 36 kDa
• UniProt ID: P29965
• 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 TRAP/CD40L/CD40LG using anti-TRAP/CD40L/CD40LG antibody (orb745937). 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 30ug of sample under reducing conditions. Lane 1: human HL-60 whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human Raji whole cell lysates, Lane 4: human Jurkat whole cell lysates, Lane 5: rat RH35 whole cell lysates, Lane 6: mouse spleen tissue lysates, Lane 7: mouse ANA-1 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-TRAP/CD40L/CD40LG antigen affinity purified polyclonal antibody (Catalog # orb745937) 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-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 TRAP/CD40L/CD40LG at approximately 36KD. The expected band size for TRAP/CD40L/CD40LG is at 36KD.

Flow Cytometry analysis of C6 cells using anti-TRAP/CD40L/CD40LG antibody (orb745937). Overlay histogram showing C6 cells stained with orb745937 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRAP/CD40L/CD40LG Antibody (orb745937, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Flow Cytometry analysis of human PBMC cells using anti-TRAP/CD40L/CD40LG antibody (orb745937). Overlay histogram showing human PBMC cells stained with orb745937 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRAP/CD40L/CD40LG Antibody (orb745937, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.