Anti gp130 antibody

Manufactured by Merck Group

The Anti-GP130 antibody is a research tool used in laboratory settings. It binds to the GP130 protein, which is a component of various cytokine receptors. The primary function of this antibody is to facilitate the study of GP130-related signaling pathways and their involvement in biological processes.

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Lab products found in correlation

Protein a g agarose, by Thermo Fisher Scientific (2 mentions) Ip buffer, by Thermo Fisher Scientific (2 mentions) Lysis buffer, by Cell Signaling Technology (1 mentions) Anti mouse igg hrp linked secondary antibody, by Cell Signaling Technology (1 mentions)

2 protocols using anti gp130 antibody

Investigating GP130 Signaling Pathway in RD Sarcoma Cells

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RD sarcoma cells were treated with Bazedoxifene 15μM for 16 hours. Cell lysates were prepared by adding lysis buffer (Cell Signaling Technology, Danvers, MA) and were pre-cleared using protein A/G agarose (Pierce Biotechnology, Rockford, IL), incubating with gentle mixing at 4°C for 2 h. GP130 was immunoprecipitated by incubating cell lysates with anti-GP130 antibody (EMD Millipore Corporation, Temecula, CA) overnight at 4°C. Protein agarose slurry was added and further incubated for 2 h at 4°C. At the end of incubation, protein A/G agarose was washed three times with IP buffer (Thermo, #28379) and proteins bound to GP130 were collected by boiling the samples in 5x loading buffer. The supernant was then separated by 10% SDS-PAGE and subjected to western blot analysis with 1:1000 dilution of primary antibodies and 1:10000 horseradish peroxidase-conjugated secondary antibodies. Antibodies against IL-6R, STAT3, GP130, and JAK1 were used for western blotting.

Xiao H., Bid H.K., Chen X., Wu X., Wei J., Bian Y., Zhao C., Li H., Li C, & Lin J. (2017). Repositioning Bazedoxifene as a novel IL-6/GP130 signaling antagonist for human rhabdomyosarcoma therapy. PLoS ONE, 12(7), e0180297.

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Bazedoxifene Modulates IL-6 Signaling

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Cells were pretreated with bazedoxifene (20 μM) in serum free medium for 4 hours followed by adding IL-6 (50 ng/mL). After additional incubation for 30 minutes and 6 hours respectively, cells were harvested and lysed. Cell lysates were precleared using protein A/G agarose (Pierce Biotechnology, Rockford, IL), incubating with gentle mixing at 4 °C for 2 hours. GP130 was immunoprecipitated by incubating cell lysates with anti-GP130 antibody (EMD Millipore Corporation, Temecula, CA) overnight at 4 °C. Protein agarose slurry was added and further incubated for 2 hours at 4 °C. At the end of incubation, protein A/G agarose was washed three times with IP buffer (Thermo, #28379) and proteins bound to GP130 were collected by boiling the samples in SDS loading buffer. Supernatant was then separated by 10% SDS-PAGE and subjected to western blotting analysis. Monoclonal human IL-6Rα mouse antibody, phosphorylated STAT3 (Y705) rabbit antibody, STAT3 rabbit antibody, GP130 rabbit antibody and the anti-mouse IgG, HRP-linked secondary antibody were purchased from Cell Signaling Technology. JAK1 mouse primary antibody was obtained from R&D System (#MAB42601).

Chen X., Tian J., Su G.H, & Lin J. (2019). Blocking IL-6/GP130 Signaling Inhibits Cell Viability/Proliferation, Glycolysis, and Colony Forming Activity in Human Pancreatic Cancer Cells. Current cancer drug targets, 19(5), 417-427.

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