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Product details for SOCS-3-Biotin antibody
SOCS-3-Biotin antibody
Overview
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| Antigen: | SOCS-3-Biotin |
| Antibody Type: | Monoclonal |
| Application: | Western Blotting (WB) |
| Host: | Mouse |
| Quantity: | |
| Price: | 266,00 € (Plus shipping costs and VAT) |
| Order number: | ABIN131761 |
| Availability: | 7 to 10 days until shipment |
Additional Information: SOCS-3-Biotin antibody
| Antigen | SOCS-3-Biotin |
| 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 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 (30 kDa) on Western blotting. |
Application Details: SOCS-3-Biotin antibody
| Application | Western Blotting (WB) |
| 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 10µg of Anti-SOCS-3 monoclonal antibody (MBL, code no. K0177-3) 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. 6) Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 7) 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. 8) 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. 9) Incubate the membrane with 5 µg/mL of Biotin Labeled anti-SOCS-3 monoclonal antibody (MBL, code no. Western blotting: 5 µg/mL for chemiluminescence detection systems. This antibody detects recombinant GST-SOCS-3 but did not detect endogenous SOCS-3 in the cell lines that MBL tested. Immunoprecipitation: not tested: the unlabeled antibody. has been used successfully at 10 µg / 200 µL of cell extract from 5x10 6 cells. Immunohistochemistry: Not tested.. Immunocytochemistry: Not tested. Flow Cytometry: Not tested. |
| Concentration | 20µg/mL |
| Buffer | 20µg IgG in 1mL volume of PBS containing 1% BSA and 0.1% NaN 3 . *Azide may react with copper or lead in plumbing system to form explosive metal azides. Therefore, always flush drains with plenty of water when of disposing materials containing azide. |
| Research Area | Signaling, Cytokines |
| Restrictions | For Research Use Only |
Publications: SOCS-3-Biotin 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: SOCS-3-Biotin antibody
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Immunoprecipitation of SOCS-3 from mouse hepatocytes with K0177-3 (1) or mouse IgG1 (2). After immunoprecipitated with the antibody, immunocomplex was resolved on SDS-PAGE and immunoblotted with ABIN131761. |
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