The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Use a concentration of 1 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 24.7 kDa).Can be blocked with Human SOCS3 peptide (ab16199).
Use at an assay dependent concentration. PubMed: 19855398
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
SOCS family proteins form part of a classical negative feedback system that regulates cytokine signal transduction. SOCS3 is involved in negative regulation of cytokines that signal through the JAK/STAT pathway. Inhibits cytokine signal transduction by binding to tyrosine kinase receptors including gp130, LIF, erythropoietin, insulin, IL12, GCSF and leptin receptors. Binding to JAK2 inhibits its kinase activity. Suppresses fetal liver erythropoiesis. Regulates onset and maintenance of allergic responses mediated by T-helper type 2 cells. Regulates IL-6 signaling in vivo (By similarity). Probable substrate recognition component of a SCF-like ECS (Elongin BC-CUL2/5-SOCS-box protein) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins. Seems to recognize IL6ST.
Widely expressed with high expression in heart, placenta, skeletal muscle, peripheral blood leukocytes, fetal and adult lung, and fetal liver and kidney. Lower levels in thymus.
Protein modification; protein ubiquitination.
Involvement in disease
Note=There is some evidence that SOCS3 may be a susceptibility gene for atopic dermatitis linked to 17q25. SOCS3 messenger RNA is significantly more highly expressed in skin from patients with atopic dermatitis than in skin from healthy controls. Furthermore, a genetic association between atopic dermatitis and a haplotype in the SOCS3 gene has been found in two independent groups of patients.
The ESS and SH2 domains are required for JAK phosphotyrosine binding. Further interaction with the KIR domain is necessary for signal and kinase inhibition. The SOCS box domain mediates the interaction with the Elongin BC complex, an adapter module in different E3 ubiquitin ligase complexes.
Phosphorylated on tyrosine residues after stimulation by the cytokines, IL-2, EPO or IGF1.
ICC/IF image of ab16030 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab16030, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
ab16030 staining SOCS3 in human bone marrow showing strong staining within the bone marrow. Paraffin embedded human bone marrow was incubated with ab16030 (1/100) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH6.
ab16030 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Immunohistochemistry (Frozen sections) - Anti-SOCS3 antibody (ab16030)This image is courtesy of an anonymous Abreview
ab16030 staining SOCS3 in Human pancreatic tumour tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/300 in PBS) for 8 hours at 4°C. An Alexa Fluor®555-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. Nuclei were stained by DAPI.
Western blot - Anti-SOCS3 antibody (ab16030)This image is courtesy of an anonymous Abreview
All lanes : Anti-SOCS3 antibody (ab16030) at 1 µg/ml
Lane 1 : Untreated Human peripheral blood mononuclear cell lysate Lane 2 : Human peripheral blood mononuclear cell lysate (treated with aCD3/28) Lane 3 : Human peripheral blood mononuclear cell lysate (treated with IFNγ) Lane 4 : SOCS3 protein
Lysates/proteins at 15 µg per lane.
Secondary All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG at 1/2000 dilution
Hwang M et al. Epac2a-null mice exhibit obesity-prone nature more susceptible to leptin resistance. Int J Obes (Lond)41:279-288 (2017).
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Zheng XB et al. miR-203 inhibits augmented proliferation and metastasis of hepatocellular carcinoma residual in the promoted regenerating liver. Cancer Sci108:338-346 (2017).
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