TRIM16 Antibody

• Catalog Number: orb745932
• Category: Antibodies
• Description: TRIM16 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, IHC, WB
• Reactivity: Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived mouse TRIM16 recombinant protein (Position: D65-K210).
• 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, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5μg/ml, Mouse Direct ELISA, 0.1-0.5μg/ml, Mouse
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 63 kDa
• UniProt ID: Q99PP9
• 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 TRIM16 using anti-TRIM16 antibody (orb745932). 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: rat testis tissue lysates, Lane 2: rat C6 whole cell lysates, Lane 3: mouse NIH/3T3 whole cell lysates, Lane 4: mouse Neuro-2a 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-TRIM16 antigen affinity purified polyclonal antibody (Catalog # orb745932) 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 TRIM16 at approximately 63KD. The expected band size for TRIM16 is at 63KD.

IHC analysis of TRIM16 using anti-TRIM16 antibody (orb745932). TRIM16 was detected in paraffin-embedded section of mouse intestine 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 2μg/ml rabbit anti-TRIM16 Antibody (orb745932) 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) (Catalog # orb90444) with DAB as the chromogen.

IHC analysis of TRIM16 using anti-TRIM16 antibody (orb745932). TRIM16 was detected in paraffin-embedded section of rat intestine 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 2μg/ml rabbit anti-TRIM16 Antibody (orb745932) 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) (Catalog # orb90444) with DAB as the chromogen.

IF analysis of TRIM16 using anti-TRIM16 antibody (orb745932). TRIM16 was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-TRIM16 Antibody (orb745932) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.