Anti mouse and anti rabbit secondary antibodies conjugated to hrp

Manufactured by Cell Signaling Technology

Anti-mouse and anti-rabbit secondary antibodies conjugated to HRP are used to detect the presence of primary antibodies in various immunoassays. These secondary antibodies bind to the constant region of the primary antibodies and are conjugated to the enzyme horseradish peroxidase (HRP), which can be used to generate a colorimetric or chemiluminescent signal for visualization and quantification.

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Lab products found in correlation

Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (2 mentions) Dab kit, by Cell Signaling Technology (2 mentions) Hematoxylin, by Vector Laboratories (2 mentions) Bolt bis tris plus gel, by Thermo Fisher Scientific (2 mentions) Pierce ecl western blotting substrate, by Bio-Rad (2 mentions) Ripa buffer, by Cell Signaling Technology (2 mentions) Restore plus western blot stripping buffer, by Thermo Fisher Scientific (2 mentions) Chemidoc plus imaging system, by Bio-Rad (2 mentions) Ab32081, by Abcam (2 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Immobilon fl pvdf membrane, by Merck Group (2 mentions) Protein assay, by Bio-Rad (2 mentions) F4 80, by Thermo Fisher Scientific (1 mentions) Ethylene diamine tetraacetic acid (edta), by Cell Signaling Technology (1 mentions) Citrate buffer, by Vector Laboratories (1 mentions) γ h2ax, by Abcam (1 mentions) Sodium citrate buffer, by Vector Laboratories (1 mentions) Hpa021241, by Abcam (1 mentions) Ab8226, by Abcam (1 mentions)

Spelling variants of this product

Anti-mouse and anti-rabbit secondary antibodies (33 citations) Anti-rabbit and anti-mouse HRP conjugated secondary antibodies (29 citations) HRP-conjugated antibodies (17 citations) Anti-mouse secondary antibodies (6 citations) Secondary antibodies conjugated to HRP (5 citations) Anti-rabbit HRP secondary antibodies (3 citations) HRP-conjugated anti-mouse and anti-rabbit antibodies (2 citations) Secondary HRP-conjugated anti-rabbit (2 citations) Anti-mouse/rabbit secondary antibodies (2 citations) Anti-rabbit and anti-mouse antibodies (2 citations)

4 protocols using anti mouse and anti rabbit secondary antibodies conjugated to hrp

Molecular Profiling of Breast Tumor Microenvironment

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Brca1^Co/Co;MMTV-Cre;p53^+/- mice were treated with five doses (three times a week) of saline, i.v.TLZ or NanoTLZ (N=5/group). Tumor, mammary gland and spleen were then harvested and sectioned for histopathology and immunohistochemistry. EDTA (Cell Signaling, for CD3) or citrate buffer (Vector, Cat. # H3300, for all the other antibodies) was used for antigen retrieval. Endogenous peroxidase activity was quenched using hydrogen peroxide (3%) for 10 minutes. Sections were stained with CD45 (1:100, BioScience), CD3 (1:40, Biolegend), Gr-1 (1:50, R&D), F4/80 (1:50, Invitrogen), Foxp3 (1:25, BioScience), PCNA (1:200, Santa Cruz), or γH₂AX (1:100, Abcam) antibodies. Anti-rat secondary antibody was purchased from Vector. Anti-mouse and anti-rabbit secondary antibodies conjugated to HRP were purchased from Cell Signaling. Signal was detected using a DAB kit (Cell Signaling). Sections were counterstained with hematoxylin (Vector).

Zhang D., Baldwin P., Leal A.S., Carapellucci S., Sridhar S, & Liby K.T. (2019). A nano-liposome formulation of the PARP inhibitor Talazoparib enhances treatment efficacy and modulates immune cell populations in mammary tumors of BRCA-deficient mice. Theranostics, 9(21), 6224-6238.

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Western Blot Analysis of Cellular Proteins

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Tissue and cell lysates were prepared using RIPA buffer (Cell Signaling Technologies) supplemented with protease inhibitor cocktail (Thermo Fisher Scientific). Protein concentrations were quantitated using BioRad Protein Assay. Equal amounts of protein (10-30μg) were separated on 4-12% Bolt bis-tris plus gels (Thermo Fisher Scientific) and transferred to Immobilon FL PVDF membranes (Millipore Sigma), which were blocked in 5% milk in TBS-T at room temperature for 1 hour and incubated with primary antibodies at 4°C overnight. Blots were incubated with anti-mouse and anti-rabbit secondary antibodies conjugated to HRP (Cell Signaling Technologies) at room temperature for 1 hour before visualization with Pierce ECL Western Blotting Substrate in a BioRad ChemiDoc Plus Imaging system. Blots were stripped with RestorePlus Western Blot Stripping Buffer (Thermo) prior to probing for the loading control.
The following primary antibodies were used: PHGDH (rabbit polyclonal, Sigma #HPA021241; rabbit mAb, Cell Signaling Technologies – CST #66350), ERK2 (rabbit mAb, Abcam #ab32081); β-actin (mouse mAb, Abcam #ab8226; rabbit mAb, CST #8457); HSP90 (rabbit polyclonal, CST #4874); Rad50 Antibody (Rabbit polyclonal, CST #3427), Phospho-Rad50 (Ser635) (Rabbit polyclonal, CST #14223), Phospho-ATM (Ser1981) (D6H9, Rabbit mAb, CST #5883), ATM (D2E2, Rabbit mAb, CST #2873).

Ngo B., Kim E., Osorio-Vasquez V., Doll S., Bustraan S., Liang R.J., Luengo A., Davidson S.M., Ali A., Ferraro G.B., Fischer G.M., Eskandari R., Kang D.S., Ni J., Plasger A., Rajasekhar V.K., Kastenhuber E.R., Bacha S., Sriram R.K., Stein B.D., Bakhoum S.F., Snuderl M., Cotzia P., Healey J.H., Mainolfi N., Suri V., Friedman A., Manfredi M., Sabatini D.M., Jones D.R., Yu M., Zhao J.J., Jain R.K., Keshari K.R., Davies M.A., Vander Heiden M.G., Hernando E., Mann M., Cantley L.C, & Pacold M.E. (2020). Limited Environmental Serine and Glycine Confer Brain Metastasis Sensitivity to PHGDH Inhibition. Cancer discovery, 10(9), 1352-1373.

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Immunohistochemical Staining of Tumors

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Tumors were harvested and fixed in 10% neutral buffered formalin. Sections were obtained for immunohistochemistry staining. Sodium citrate buffer (10 mM, Vector) was used for antigen retrieval and 3% hydrogen peroxide (15-minute incubation) was used to quench the endogenous peroxidase activity. Sections were stained with PCNA (1:200, Santa Cruz) or Nrf2 (1:200, Novus Biologicals) antibodies for 1 hour at room temperature or overnight at 4°C, respectively. Anti-mouse and anti-rabbit secondary antibodies conjugated to HRP were purchased from Cell Signaling Technology. Signal was detected using a DAB kit (Cell Signaling Technology) and sections were counterstained with hematoxylin (Vector).

Zhang D., Hou Z., Aldrich K.E., Lockwood L., Odom A.L, & Liby K.T. (2021). A novel Nrf2 pathway inhibitor sensitizes Keap1-mutant lung cancer cells to chemotherapy. Molecular cancer therapeutics, 20(9), 1692-1701.

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Western Blot Protein Expression Analysis

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Tissue and cell lysates were prepared using RIPA buffer (Cell Signaling Technologies) supplemented with protease inhibitor cocktail (Thermo Fisher Scientific). Protein concentrations were quantitated using BioRad Protein Assay. Equal amounts of protein (10-30 µg) were separated on 4-12% Bolt bis-tris plus gels (Thermo Fisher Scientific) and transferred to Immobilon FL PVDF membranes (Millipore Sigma), which were blocked in 5% milk in TBS-T at room temperature for 1 hour and incubated with primary antibodies at 4 °C overnight. Blots were incubated with anti-mouse and anti-rabbit secondary antibodies conjugated to HRP (Cell Signaling Technologies) at room temperature for 1 hour before visualization with Pierce ECL Western Blotting Substrate in a BioRad ChemiDoc Plus Imaging system. Blots were stripped with RestorePlus Western Blot Stripping Buffer (Thermo) prior to probing for the loading control. The following primary antibodies were used: PHGDH (Rabbit Polyclonal; Sigma HPA021241), ERK2 (Rabbit Monoclonal; AbCam ab32081); b-actin (Mouse Monoclonal; Abcam ab8226); HSP90 (Rabbit Polyclonal, Cell Signaling Technologies 4874).

Ngo B., Kim E.D., Osorio-Vasquez V., Doll S., Bustraan S., Luengo A., Davidson S.M., Ali A., Ferraro G.D., Kang D.S., Ni J., Liang R.J., Plasger A., Kastenhuber E.R., Eskandari R., Bacha S., Siriam R.K., Bakhoum S.F., Mullarky E., Snuderl M., Cotzia P., Mainolfi N., Suri V., Friedman A., Manfredi M., Sabatini D.M., Jones D.R., Yu M., Zhao J.J., Jain R.K., Heiden M.G.V., Hernando E., Mann M., Cantley L.C., & Pacold M.E. (2020). Limited Environmental Serine Confers Sensitivity to PHGDH Inhibition in Brain Metastasis.

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