Anti-Filamin A/FLNA Antibody (monoclonal, 3F8)

• Catalog Number: orb692222
• Category: Antibodies
• Description: Anti-Filamin A/FLNA Antibody (monoclonal, 3F8). Tested in IF, IHC, WB applications. This antibody reacts with Human.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG1
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human Filamin A/FLNA recombinant protein (Position: E451-Q638).
• UniProt ID: P21333
• MW: 281 kDa
• Tested applications: IF, IHC, WB
• Application notes: Western blot, 0.25-0.5μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human Immunofluorescence, 5μg/ml, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Antibody Type: Primary Antibody
• Clone Number: 3F8
• 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.
• Alternative names: T-cell surface glycoprotein CD5; Lymphocyte antige
• Note: For research use only

IF analysis of Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human rectal cancer 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 5 µg/mL mouse anti-Filamin A/FLNA Antibody overnight at 4°C. Biotin conjugated goat anti-mouse IgG 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 Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human adrenocortical adenoma 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 mouse anti-Filamin A/FLNA Antibody 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) with DAB as the chromogen.

IHC analysis of Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human appendicitis 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 mouse anti-Filamin A/FLNA Antibody 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) with DAB as the chromogen.

IHC analysis of Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human breast cancer 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 mouse anti-Filamin A/FLNA Antibody 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) with DAB as the chromogen.

IHC analysis of Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human esophageal squamous carcinomar 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 mouse anti-Filamin A/FLNA Antibody 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) with DAB as the chromogen.

IHC analysis of Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human melanoma 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 mouse anti-Filamin A/FLNA Antibody 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) with DAB as the chromogen.

IHC analysis of Filamin A/FLNA using anti-Filamin A/FLNA antibody. Filamin A/FLNA was detected in paraffin-embedded section of human rectal cancer 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 mouse anti-Filamin A/FLNA Antibody 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) with DAB as the chromogen.

Western blot analysis of Filamin A/FLNA using anti-Filamin A/FLNA 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: human A549 whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human Sw620 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 A/FLNA antigen affinity purified monoclonal 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-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 with Tanon 5200 system. A specific band was detected for Filamin A/FLNA at approximately 281 KD. The expected band size for Filamin A/FLNA is at 281 KD.