Tspan1 Antibody

• Catalog Number: orb692214
• Category: Antibodies
• Description: Tspan1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, IF, IHC, WB
• Reactivity: Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived rat Tspan1 recombinant protein (Position: A125-A212).
• 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, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Rat Immunofluorescence, 5 μg/ml, Rat Flow Cytometry, 1-3μg/1x106 cells, Rat Direct ELISA, 0.1-0.5μg/ml, Rat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 28-30 kDa
• UniProt ID: Q6AYR9
• 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 500μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Tspan1 using anti-Tspan1 antibody (orb692214). 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: rat small intestine 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 rabbit anti-Tspan1 antigen affinity purified polyclonal antibody (Catalog # orb692214) 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 Tspan1 at approximately 28-30KD. The expected band size for Tspan1 is at 28-30KD.

Flow Cytometry analysis of RH35 cells using anti-Tspan1 antibody (orb692214). Overlay histogram showing RH35 cells stained with orb692214 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Tspan1 Antibody (orb692214, 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.

IF analysis of Tspan1 using anti-Tspan1 antibody (orb692214). Tspan1 was detected in a paraffin-embedded section of rat stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-Tspan1 Antibody (orb692214) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (orb1474935) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®488 Conjugated Avidin. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Tspan1 using anti-Tspan1 antibody (orb692214). Tspan1 was detected in a paraffin-embedded section of rat stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Tspan1 Antibody (orb692214) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.