Tigit Antibody

SKU: orb692212

Description

Anti-Tigit Antibody. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Research Area

Metabolism Research, Signal Transduction

Images & Validation

• Tested Applications: ELISA, FC, IHC, WB
• Dilution range: Western blot, 0.25-0.5μg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Mouse ELISA, 0.1-0.5μg/ml, -
• Reactivity: Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived mouse Tigit recombinant protein (Position: T29-G249).
• Target: T-cell immunoreceptor with Ig and ITIM domains
• Molecular Weight: 30 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.005mg NaN3.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Insulin-like growth factor-binding protein 5; IBP-5; IGF-binding protein 5; IGFBP-5; IGFBP5; IBP5

Flow Cytometry analysis of RAW264.7 cells using anti-Tigit antibody. Overlay histogram showing RAW264.7 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Tigit 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.

IHC analysis of Tigit using anti-Tigit antibody. Tigit was detected in paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-Tigit Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of Tigit using anti-Tigit 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 50 ug of sample under reducing conditions. Lane 1: rat spleen tissue lysates, Lane 2: rat plasma lysates, Lane 3: mouse spleen 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 rabbit anti-Tigit 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 Tigit at approximately 30 KD. The expected band size for Tigit is at 30 KD.

Quick Database Links

Gene Symbol

T-cell immunoreceptor with Ig and ITIM domains

UniProt

P86176