Filamin B/FLNB Antibody (monoclonal, 11E2D2)

• Catalog Number: orb865673
• Category: Antibodies
• Description: Filamin B/FLNB Antibody (monoclonal, 11E2D2)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 11E2D2
• Tested applications: ICC, IF, IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG1
• Immunogen: E.coli-derived human Filamin B/FLNB recombinant protein (Position: Q397-D701).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5 μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 278 kDa
• UniProt ID: O75369
• Storage: 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 freezing and thawing.
• 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. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Filamin B/FLNB using anti-Filamin B/FLNB antibody (orb865673). 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 A549 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 mouse anti-Filamin B/FLNB antigen affinity purified monoclonal antibody (Catalog # orb865673) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for Filamin B/FLNB at approximately 278 kDa. The expected band size for Filamin B/FLNB is at 278 kDa.

IHC analysis of Filamin B/FLNB using anti-Filamin B/FLNB antibody (orb865673). Filamin B/FLNB was detected in a paraffin-embedded section of human lung cancer 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 mouse anti-Filamin B/FLNB Antibody (orb865673) overnight at 4°C. Biotinylated goat anti-mouse 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 # orb90443) with DAB as the chromogen.

IHC analysis of Filamin B/FLNB using anti-Filamin B/FLNB antibody (orb865673). Filamin B/FLNB was detected in a paraffin-embedded section of human lymph node of rectal cancer 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 mouse anti-Filamin B/FLNB Antibody (orb865673) overnight at 4°C. Biotinylated goat anti-mouse 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 # orb90443) with DAB as the chromogen.

IHC analysis of Filamin B/FLNB using anti-Filamin B/FLNB antibody (orb865673). Filamin B/FLNB was detected in a paraffin-embedded section of human metaplasia of squamous cells of the renal pelvis 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 mouse anti-Filamin B/FLNB Antibody (orb865673) overnight at 4°C. Biotinylated goat anti-mouse 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 # orb90443) with DAB as the chromogen.

IHC analysis of Filamin B/FLNB using anti-Filamin B/FLNB antibody (orb865673). Filamin B/FLNB was detected in a paraffin-embedded section of human spleen 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 mouse anti-Filamin B/FLNB Antibody (orb865673) overnight at 4°C. Biotinylated goat anti-mouse 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 # orb90443) with DAB as the chromogen.

IF analysis of Filamin B/FLNB using anti-Filamin B/FLNB antibody (orb865673). Filamin B/FLNB was detected in an immunocytochemical section of HeLa 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 mouse anti-Filamin B/FLNB Antibody (orb865673) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse 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.