Gelsolin (also known as brevin, Actin-depolymerizing factor or ADF), a protein of leukocytes, platelets and other cells, severs Actin filaments in the presence of submicromolar calcium, thereby isolating cytoplasmic Actin gels. A calcium-independent mechanism reverses the process. A Gelsolin variant with 23 more amino-terminal amino acids is a plasma component probably involved in the clearance of Actin, the most abundant human protein, from the circulation. It has been suggested that a single gene encodes both cell and plasma gelsolins. Gelsolin may be unique in that it is made for both secretion and intracytoplasmic location. Amino acid homology was identified between Gelsolin and the amyloid of the Finnish variety of amyloidosis. The amyloid in this disorder is antigenically and structurally related to Gelsolin. Gelsolin is the principal intracellular and extracellular Actin-severing protein. Gelsolin and Gc protein together constitute the extracellular Actin-scavenger system which prevents the toxic effects of Actin release into the extracellular space under circumstances of cell necrosis.
Background References
1. Kim J et al. Functional genomic screen for modulators of ciliogenesis and cilium length. Nature 464:1048-1051 (2010).
2. Ghiso J et al. Gelsolin variant (Asn-187) in familial amyloidosis, Finnish type. Biochem J 272:827-830 (1990).
Western blot analysis of Gelsolin on different lysates with Rabbit anti-Gelsolin antibody (ET7107-45) at 1/1,000 dilution.
Lane 1: THP-1 cell lysate (20 µg/Lane) Lane 2: HepG2 cell lysate (20 µg/Lane) Lane 3: NIH/3T3 cell lysate (20 µg/Lane) Lane 4: COS-1 cell lysate (20 µg/Lane) Lane 5: Mouse lung tissue lysate (40 µg/Lane)
Predicted band size: 86 kDa Observed band size: 86 kDa
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-45) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of Gelsolin on different lysates with Rabbit anti-Gelsolin antibody (ET7107-45) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-Gelsolin KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 86 kDa Observed band size: 86 kDa
Exposure time: 180 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-45) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Gelsolin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-45, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Gelsolin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-45, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Gelsolin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-45, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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