Anti-TRAP/CD40L/CD40LG Antibody

• Catalog Number: orb745937
• Category: Antibodies
• Description: Anti-TRAP/CD40L/CD40LG Antibody. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat.
• 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.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 36 kDa
• UniProt ID: P29965
• 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: CD40 ligand; CD40-L; T-cell antigen Gp39; TNF-rela
• Note: For research use only
• Application notes: Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human, Rat ELISA, 0.1-0.5μg/ml, -. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of C6 cells using anti-TRAP/CD40L/CD40LG antibody. Overlay histogram showing C6 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-TRAP/CD40L/CD40LG Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of human PBMC cells using anti-TRAP/CD40L/CD40LG antibody. Overlay histogram showing human PBMC cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-TRAP/CD40L/CD40LG Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of TRAP/CD40L/CD40LG using anti-TRAP/CD40L/CD40LG 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 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 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 rabbit anti-TRAP/CD40L/CD40LG antigen affinity purified polyclonal 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-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 with Tanon 5200 system. A specific band was detected for TRAP/CD40L/CD40LG at approximately 36 KD. The expected band size for TRAP/CD40L/CD40LG is at 36 KD.