Mouse igg isotype control

Manufactured by Cell Signaling Technology

Sourced in United States

The Mouse IgG Isotype Control is a laboratory reagent used to establish the appropriate level of nonspecific binding in immunoassays involving mouse immunoglobulin G (IgG). It serves as a control to differentiate specific from nonspecific antibody binding.

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Lab products found in correlation

Rabbit igg isotype control, by Cell Signaling Technology (4 mentions) Igepal ca 630, by Merck Group (2 mentions) Aprotinin, by Merck Group (2 mentions) Leupeptin, by Merck Group (2 mentions) Anti usp21, by Thermo Fisher Scientific (1 mentions) Anti stat3, by Cell Signaling Technology (1 mentions) Anti myc, by Merck Group (1 mentions) Anti phospho stat3, by Cell Signaling Technology (1 mentions) Mg132, by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions) H3k4me1, by Abcam (1 mentions) H3k4me3, by Merck Group (1 mentions) H3k27me3, by Merck Group (1 mentions) Flag tag (flag), by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) H3k27ac, by Abcam (1 mentions) Pierce crosslink immunoprecipitation kit, by Thermo Fisher Scientific (1 mentions) Lipofectamine messengermax, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Control IgG (65 citations) IgG isotype control (18 citations) Isotype IgG (12 citations) Isotype control (10 citations) Control mouse IgG (6 citations) IgG (Mouse) (3 citations)

5 protocols using mouse igg isotype control

Comprehensive Immunoblotting Protocol

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The following reagents and antibodies were used in this study:
MG132 (Sigma-Aldrich, Cat#M7449); anti-Flag (Sigma-Aldrich, Cat#F7425); anti-MYC (Sigma-Aldrich, Cat#M4439); anti-STAT3 (Cell Signaling Technology, Cat#9319S); anti-phospho-STAT3 (Cell Signaling Technology, Cat#9145S).
Mouse IgG Isotype control (Cell Signaling Technology, Cat#53484).
Rabbit IgG Isotype control (Cell Signaling Technology, Cat#2729); anti-Usp21 (Invitrogen, Cat#PA5-110556); anti-beta actin (2D4H5) (Proteintech, Cat#66 009-1-Ig).

Deng B., Yang B., Chen J., Wang S., Zhang W., Guo Y., Han Y., Li H., Dang Y., Yuan Y., Dai X., Zang Y., Li Y, & Li B. (2022). Gallic acid induces T-helper-1-like Treg cells and strengthens immune checkpoint blockade efficacy. Journal for Immunotherapy of Cancer, 10(7), e004037.

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Antibody Characterization for Chromatin Immunoprecipitation

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Antibodies used were as follows: ICN1 (Cell Signaling Technology, 2421), Rbpj (5313, Cell Signaling Technology), FLAG (F1804, Sigma), HA (3725, Cell Signaling Technology), β-actin (A5316, Sigma), MYC (D84C2; Cell Signaling Technology), and ZMIZ1 (AP6236a, Abgent). Antibodies for ChIP-Seq are as follows: H3K4me3 (Millipore, 07-473); H3K4me1 (Abcam, ab8895); H3K27me3 (Millipore #07-449); H3K27ac (Abcam, ab4729); HA (Abcam, ab9110, 2.5ug); mouse IgG isotype control (Cell Signaling Technology, 5315s); rabbit IgG isotype control (Cell Signaling Technology, 2729); Rbpj (Cell Signaling Technology, 5313); ICN1 (Cell Signaling Technology, 4147s).

Pinnell N., Yan R., Cho H.J., Keeley T., Murai M.J., Liu Y., Alarcon A.S., Qin J., Wang Q., Kuick R., Elenitoba-Johnson K.S., Maillard I., Samuelson L.C., Cierpicki T, & Chiang M.Y. (2015). The PIAS-like coactivator Zmiz1 is a direct and selective cofactor of Notch1 in T-cell development and leukemia. Immunity, 43(5), 870-883.

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Immunoprecipitation and Immunoblotting Protocol

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LNCaP cells were lysed with IGEPAL CA-630 buffer (50 mM Tris-HCl, pH 7.4, (Sigma, T5030), 1% IGEPAL CA-630 (Sigma, I8896), 10 mM EDTA, 150 mM NaCl, 50 mM NaF, 1 μM leupeptin (Sigma, L5793), and 0.1 μM aprotinin (Sigma, SRE0050)). Primary antibody was covalently immobilized on protein A/G agarose using the Pierce Crosslink Immunoprecipitation Kit according to the manufacturer’s instructions (Thermo Scientific, 26147). Samples were incubated with immobilized antibody beads for at least 2 h at 4 °C. Cell lysates were also subjected to immunoprecipitation with either mouse IgG isotype control (Cell Signaling Technology, 5415) or rabbit IgG isotype control (Cell Signaling Technology, 3900), depending on the immunoglobulin type of primary antibody. After immunoprecipitation, the samples were washed with TBS five times. They were then eluted with glycine-HCl (0.1 M, pH 3.5) and the immunoprecipitates were subjected to immunoblotting using specific primary antibodies.

Chen Z., Cai A., Zheng H., Huang H., Sun R., Cui X., Ye W., Yao Q., Chen R, & Kou L. (2020). Carbidopa suppresses prostate cancer via aryl hydrocarbon receptor-mediated ubiquitination and degradation of androgen receptor. Oncogenesis, 9(5), 49.

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Investigating TXNDC5 and TGFBR1 Interactions

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Human WT TXNDC5, HA-tagged mutant TXNDC5-AAA and human TGFBR1-Myc-Flag-tagged mRNA were transfected into NIH-3T3 fibroblasts using Lipofectamine MessengerMAX (Thermo Fisher Scientific, MA, USA) for 4 h according to manufacturer’s instructions. Protein lysates from NIH-3T3 were incubated with mouse anti-Flag tag monoclonal antibody (Cell Signaling Technology, MA,USA, #8146) or mouse IgG isotype control (Cell Signaling Technology, MA,USA, #5415 S) with protein G magnetic beads and collected with a magnetic stand. IP of WT or mutant TXNDC5 was conducted using a magnetic IP kit (Thermo Fisher Scientific, MA, USA). Proteins co-immunoprecipitated with Flag-tag, TXNDC5, or HA were eluted and subjected to gel electrophoresis and immunoblotting using the antibodies described above.

Lee T.H., Yeh C.F., Lee Y.T., Shih Y.C., Chen Y.T., Hung C.T., You M.Y., Wu P.C., Shentu T.P., Huang R.T., Lin Y.S., Wu Y.F., Lin S.J., Lu F.L., Tsao P.N., Lin T.H., Lo S.C., Tseng Y.S., Wu W.L., Chen C.N., Wu C.C., Lin S.L., Sperling A.I., Guzy R.D., Fang Y, & Yang K.C. (2020). Fibroblast-enriched endoplasmic reticulum protein TXNDC5 promotes pulmonary fibrosis by augmenting TGFβ signaling through TGFBR1 stabilization. Nature Communications, 11, 4254.

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Immunoprecipitation and Immunoblotting of MCF7 Cells

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MCF7 cells were lysed with IGEPAL CA-630 buffer: 50 mM Tris-HCl, pH 7.4, (Sigma, T5030), 1% IGEPAL CA-630 (Sigma, I8896), 10 mM EDTA, 150 mM NaCl, 50 mM NaF, 1 µM leupeptin (Sigma, L5793), 0.1 µM aprotinin (Sigma, SRE0050). Primary antibody was covalently immobilized on protein A/G agarose using the Pierce Crosslink Immunoprecipitation Kit (Thermo Scienti c, 26147). Samples were incubated with immobilized antibody beads for at least 2 h at 4°C. Cell lysates were also subjected to immunoprecipitation with either mouse IgG isotype control (Cell Signaling Technology, 5415) or rabbit IgG isotype control (Cell Signaling Technology, 3900) according to the immunoglobulin type of the primary antibody. After immunoprecipitation, the samples were washed with PBS for 5 times. They were then eluted with glycine-HCl (0.1 M, pH 3.5) and the immunoprecipitates were then subjected to immunoblotting using speci c primary antibodies described in the Western blot part.

Chen Z., Xia X., Chen H., Huang H., An X., Sun M., Yao Q., Kim K., Zhang H., Chu M., Chen R., Bhutia Y.D., Ganapathy V, & Kou L. (2022). Carbidopa suppresses estrogen receptor-positive breast cancer via AhR-mediated proteasomal degradation of ERα. Investigational new drugs, 40(6).

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