alpha Internexin/INA Antibody

• Catalog Number: orb763051
• Category: Antibodies
• Description: alpha Internexin/INA Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, ICC, IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human INA recombinant protein (Position: L62-D169).
• 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, Human Immunofluorescence, 5μg/ml, Rat Flow Cytometry, 1-3μg/1x106 cells, Human Direct ELISA, 0.1-0.5μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 66 kDa
• UniProt ID: Q16352
• 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 Alpha Internexin/INA using anti-Alpha Internexin/INA antibody (orb763051). 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: rat brain tissue lysates, Lane 2: mouse brain 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-Alpha Internexin/INA antigen affinity purified polyclonal antibody (Catalog # orb763051) 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 Alpha Internexin/INA at approximately 66 kDa. The expected band size for Alpha Internexin/INA is at 66 kDa.

IHC analysis of Alpha Internexin/INA using anti-Alpha Internexin/INA antibody (orb763051). Alpha Internexin/INA was detected in a paraffin-embedded section of mouse brain 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-Alpha Internexin/INA Antibody (orb763051) 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 Alpha Internexin/INA using anti-Alpha Internexin/INA antibody (orb763051). Alpha Internexin/INA was detected in a paraffin-embedded section of mouse brain 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-Alpha Internexin/INA Antibody (orb763051) 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 Alpha Internexin/INA using anti-Alpha Internexin/INA antibody (orb763051). Alpha Internexin/INA was detected in a paraffin-embedded section of rat brain 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-Alpha Internexin/INA Antibody (orb763051) 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 Alpha Internexin/INA using anti-Alpha Internexin/INA antibody (orb763051). Alpha Internexin/INA was detected in an immunocytochemical section of SiHa 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 5 μg/mL rabbit anti-Alpha Internexin/INA Antibody (orb763051) 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.

IF analysis of Alpha Internexin/INA using anti-Alpha Internexin/INA antibody (orb763051). Alpha Internexin/IN was detected in a paraffin-embedded section of rat brain 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-Alpha Internexin/INA Antibody (orb763051) 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.

Flow Cytometry analysis of A549 cells using anti-Alpha Internexin/INA antibody (orb763051). Overlay histogram showing A549 cells stained with orb763051 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Alpha Internexin/INA Antibody (orb763051, 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 (Red line) was also used as a control.