Anti-WASH1 antibody, Mouse monoclonal
SIGMA/SAB4200552 - clone WASH1-27, purified from hybridoma cell culture
Synonym: Anti-CXYorf1-like protein on chromosome 9; Anti-FAM39E; Anti-WAS protein family homolog 1; Anti-WASH; Monoclonal Anti-WASH1 antibody produced in mouse
Product Type: Chemical
Immunofluorescence WT MEF cells (WASH +/+) or WASHout MEF cells (WASH-/-) were fixed in 4% PFA for 20 minutes and permeabilized with 0.15% Surfactamps for 3 minutes. Fixed cells were stained with 1:300 dilution of Monoclonal Anti-WASH1 Clone: WASH1-27 (Cat. No. SAB4200552). The antibody was developed using Goat Anti-Mouse IgG, TRITC conjugate. Cells were counterstained with Hoechst (blue) to stain nuclei. Pictures are courtesy of Timothy S. Gomez and Daniel D. Billadeau, Departments of Biochemistry and Molecular Biology and Immunology, Division of Oncology Research, Mayo Clinic, Rochester, USA.
Immunoblotting Whole extract of HEK-293T cells overexpressing mouse WASH1 was separated on SDS-PAGE and probed with Monoclonal Anti-WASH1 Clone: WASH1-27 (Cat. No. SAB4200552). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. 2 μg/mL antibody 2. 5 μg/mL antibody 3. 5 μg/mL antibody + WASH1 immunizing peptide
Immunoblotting Whole extracts of WT MEF cells (WASH +/+) or WASHout MEF cells (WASH-/-) were separated on SDS-PAGE and probed with 1:1,000 dilution of Monoclonal Anti-WASH1 Clone: WASH1-27 (Cat. No. SAB4200552). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase and a chemiluminescent substrate. The picture is courtesy of Timothy S. Gomez and Daniel D. Billadeau, Departments of Biochemistry and Molecular Biology and Immunology, Division of Oncology Research, Mayo Clinic, Rochester, USA.
Immunoprecipitation Monoclonal Anti-WASH1 Clone: WASH1-27 (Cat. No. SAB4200552) was used to immunoprecipitate WASH1 from HeLa whole cell extract. Detection antibody: Rabbit Anti-WASH1 (Cat. No. SAB4200372). Lanes 1. 5 μg antibody 2. 10 μg antibody 3. Negative control: 10 μg non-relevant antibody
| antibody form | purified from hybridoma cell culture |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | WASH1-27, monoclonal |
| concentration | ~1 mg/mL |
| conjugate | unconjugated |
| form | buffered aqueous solution |
| isotype | IgG1 |
| mol wt | antigen ~72 kDa |
| Quality Level | 200  |
| shipped in | dry ice |
| species reactivity | mouse, human |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunoprecipitation (IP): 5-10 μg using lysates of human HeLa cells |
| western blot: 2.5-5.0 μg/mL using whole extracts of HEK-293T cells overexpressing mouse WASH1 |
| Application: | Monoclonal Anti-WASH1 antibody produced in mouse has been used in may be used in various immunochemical techniques including immunoblotting (~72 kDa), immunoprecipitation and immunofluorescence. |
| Biochem/physiol Actions: | WASH1 (Wiskott-Aldrich Syndrome Protein and SCAR Homolog), is a nucleation-promoting factor at the surface of endosomes. It recruits and activates the Arp2/3 complex to induce actin polymerization, playing a key role in the fission of endosomes. It associates with tubulin and localizes to early and recycling endosomes, where together with the Arp2/3 complex and actin, it is required for maintaining the shape of the endosomal compartment and the regulation of the retrograde transport. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | Monoclonal Anti-WASH1 (mouse IgG1 isotype) is derived from the hybridoma WASH1-27 produced by the fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with a synthetic peptide corresponding to an internal region of human WASH1. WASH1 (Wiskott-Aldrich Syndrome Protein and SCAR Homolog), a new member of the Wiskott−Aldrich Syndrome protein (WASP) family. Similar to other WASP family members, it contains a C-terminal for ′WH2, connecting and acidic′ (WCA) domain that binds to actin and the Arp2/3 complex. In addition, WASH1 also contains a short proline-rich region, a unique N-terminal domain termed WASH-homology domain (WAHD1), and a tubulin-binding region. Through its WAHD1 region, it interacts with family with sequence similarity 21 (FAM21), a protein that links WASH1 to endosomes. WASH1 forms part of a multiprotein complex composed of FAM21, KIAA1033 strumpellin and WASH-interacting protein (SWIP) and coiled coil domain containing 53 (CCDC53). |
| Immunogen: | a synthetic peptide corresponding to an internal region of human WASH1 (GeneID: 100287171), conjugated to KLH. The corresponding sequence differs by one amino acid in mouse and rat. |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |