Gelsolin/GSN Antibody

• Catalog Number: orb654260
• Category: Antibodies
• Description: Gelsolin/GSN Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human Gelsolin recombinant protein (Position: E580-A782). Human Gelsolin shares 94% and 95% amino acid (aa) sequences identity with mouse and rat Gelsolin, respectively.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat, Monkey Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 85698 MW
• UniProt ID: P06396
• 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.
• Alternative names: Gelsolin;AGEL;Actin-depolymerizing factor;ADF;Brev
• Note: For research use only
• Application notes: WB: The detection limit for Gelsolin is approximately 0.25ng/lane under reducing conditions. Tested Species: In-house tested species with positive results. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. 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.

IHC analysis of Gelsolin using anti-Gelsolin antibody (orb654260). Gelsolin was detected in a paraffin-embedded section of mouse 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 1 μg/ml rabbit anti-Gelsolin Antibody (orb654260) 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 Gelsolin using anti-Gelsolin antibody (orb654260). Gelsolin was detected in a paraffin-embedded section of rat 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 1 μg/ml rabbit anti-Gelsolin Antibody (orb654260) 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.

Western blot analysis of Gelsolin using anti-Gelsolin antibody (orb654260). 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 HeLa whole cell lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: monkey COS-7 whole cell lysates, Lane 5: monkey kidney tissue lysates, Lane 6: human CACO-2 whole cell lysates, Lane 7: rat PC-12 whole cell lysates, Lane 8: mouse RAW264.7 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 rabbit anti-Gelsolin antigen affinity purified polyclonal antibody (Catalog # orb654260) 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 Gelsolin at approximately 86 kDa. The expected band size for Gelsolin is at 86 kDa.

IHC analysis of Gelsolin using anti-Gelsolin antibody (orb654260). Gelsolin was detected in a paraffin-embedded section of human mammary 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 1 μg/ml rabbit anti-Gelsolin Antibody (orb654260) 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.