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Product details for SOCS3 antibody
SOCS3 antibody
Overview
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| Antigen: | SOCS3 |
| Antibody Type: | Monoclonal |
| Application: | Western Blotting (WB), Immunoprecipitation (IP) |
| Host: | Mouse |
| Quantity: | |
| Price: | 266,00 € (Plus shipping costs and VAT) |
| Order number: | ABIN131760 |
| Availability: | 7 to 10 days until shipment |
Additional Information: SOCS3 antibody
| Antigen | SOCS3 |
| Immunogen | This antibody was purified from hybridoma (clone 1B2) supernatant using protein A agarose. This hybridoma was established by fusion of mouse myeloma cell SP2/0 with C57BL/6 mouse splenocyte immunized with a mouse recombinant GST-SOCS-3 (2-44 a.a.). |
| Antibody Type | Monoclonal |
| Isotype | IgG1 »Matching secondary antibodies |
| Description | The Suppressor of Cytokine Signaling (SOCS) family of proteins acts in a classic negative feedback loop to regulate cytokine signal transduction. Socs genes are induced by cytokines to express SOCS proteins, which then inhibit cytokine-induced signaling. SOCS-3, also known as CIS3 or SSI3, inhibits cytokine signaling by multiple cytokines and hormones, including Epo, prolactin, growth hormone (GH), IL-2, IL-3, IL -6, IL-11, leptin, ciliary neurotrophic factor, MCSF, and LIF. The SH2 domain of SOCS-3 also interacts with the cytoplasmic domains of phosphorylated GH receptor, leptin receptor, IGF-I receptor, and the gp130 common receptor subunit. SOCS-3 directly inhibits JAK2 catalytic activity, and silencing of SOCS-3 by methylation may be an important mechanism of constitutive activation of the JAK/STAT pathway in cancer pathogenesis. |
| Clone | 1B2 |
| Host | Mouse |
| Specificity | This antibody reacts with SOCS-3. |
Application Details: SOCS3 antibody
| Application | Western Blotting (WB), Immunoprecipitation (IP) |
| Application Notes | Immunoprecipitation 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds) 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. 3) Add primary antibody as suggested in the Applications into 200 µL of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4 o C. Add 20 µL of 50% protein G agarose resuspended in the cold Lysis buffer. Mix well and incubate with gentle agitation for 60 minutes at 4 o C. 4) Wash the agarose 3-5 times with the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 5) Resuspend the agarose in 20 µL of Laemmli’s sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 6) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system (Transfer Buffer - 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for specific transfer procedure. 7) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 8) Incubate the membrane with 5 µg/mL of biotinylated anti-SOCS-3 monoclonal antibody diluted with PBS, pH 7.2 containing 1% skimmed milk for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 9) Wash the membrane with PBS (5 minutes x 6 times) 10) Incubate the membrane with the 1:20,000 POD-conjugated streptavidin (MBL, code no. IM-0309) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 11) Wash the membrane with PBS (5 minutes x 6 times). 12) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 13) Expose to X-ray film in a dark room for 5 minutes. Develop the film as usual. The conditions for exposure and development may vary. Positive control for Immunoprecipitation, mouse hepatocytes 061031-1 Western blotting: 0.5 µg/mL for chemiluminescence detection system. This antibody detects recombinant GST-SOCS-3, but does not detect SOCS-3 in the cell lines that MBL tested. Immunoprecipitation: 10 µg / 200 µL of cell extract from 5x10 6 cells. Immunohistochemistry: Not tested. Immunocytochemistry: Not tested. Flow Cytometry: Not tested. |
| Concentration | 1 mg/mL |
| Buffer | 100 µg IgG in 100 µL volume of PBS containing 50% glycerol, pH 7.2. Contains no preservatives. 0175-3 Anti-SOCS-1 (4H1) |
| Research Area | Signaling, Cytokines |
| Restrictions | For Research Use Only |
Publications: SOCS3 antibody
| Publications |
Croker, Krebs, Zhang et al.: "SOCS3 negatively regulates IL-6 signaling in vivo." In: Nature immunology , Vol. 4, Issue 6, pp. 540-5, 2003/01 (PubMed) Stoiber, Stockinger, Steinlein et al.: "Listeria monocytogenes modulates macrophage cytokine responses through STAT serine phosphorylation and the induction of suppressor of cytokine signaling 3." In: Journal of immunology (Baltimore, Md. : 1950) , Vol. 166, Issue 1, pp. 466-72, 2001/01 (PubMed) Zhang, Metcalf, Rakar et al.: "The SOCS box of suppressor of cytokine signaling-1 is important for inhibition of cytokine action in vivo." In: Proceedings of the National Academy of Sciences of the United States of America , Vol. 98, Issue 23, pp. 13261-5, 2001/01 (PubMed) |
Images: SOCS3 antibody
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Western blot analysis of GST-SOCS-3 (2-44 a.a.) using ABIN131760
Immunoprecipitation of SOCS-3 from mouse hepatocytes with ABIN131760 (1) or mouse IgG1 (2) After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoblotted with biotinylated anti-SOCS-3 monoclonal antibody. |
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