Antibody cross linking kit

Manufactured by Thermo Fisher Scientific

The Antibody Cross-linking Kit is a laboratory product designed to facilitate the covalent linkage of antibodies to solid support materials, such as beads or plates. The kit provides the necessary reagents and protocols to enable the stable attachment of antibodies, which can then be used in various immunological applications, such as affinity purification, immunoprecipitation, or enzyme-linked immunosorbent assays (ELISAs).

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Lab products found in correlation

Anti smc3, by Fortis Life Sciences (1 mentions) Anti β actin, by Merck Group (1 mentions) Horseradish peroxidase conjugated anti rabbit or anti mouse secondary antibodies, by Jackson ImmunoResearch (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Anti flag m2, by Merck Group (1 mentions) Flag m2 bead, by Merck Group (1 mentions)

2 protocols using antibody cross linking kit

Immunoprecipitation and Western Blot Analysis

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Cells were lysed in RIPA buffer containing a protease inhibitor cocktail (Roche) and MG132. Antibodies used for IP were conjugated with protein A/G beads using the antibody cross-linking kit (Pierce Biotechnology, Rockford, IL) following the manufacturer’s instructions. For western blot analysis, proteins were resolved using 10% SDS-PAGE. The antibodies used included anti-FLAG (M2; Sigma), anti-HDAC8 (H145, E5; Santa Cruz), anti-CBFβ (141,4,1; Santa Cruz), anti-p53 (DO-1, 1C12), anti-Ac-p53 (K379) (Cell Signaling, Danvers, MA), anti-SMC3 (Bethyl Laboratories), anti-Ac-SMC3 (MBL International) and anti-β-actin (Sigma, St. Louis, MO). Horseradish peroxidase–conjugated anti-rabbit or anti-mouse secondary antibodies (Jackson ImmunoResearch, West Grove PA) were used, followed by detection using the SuperFemto kit (Pierce Biotechnology, Rockford, IL).

Qi J., Singh S., Hua W.K., Cai Q., Chao S.W., Li L., Liu H., Ho Y., McDonald T., Lin A., Marcucci G., Bhatia R., Huang W.J., Chang C.I, & Kuo Y.H. (2015). HDAC8 Inhibition Specifically Targets Inv(16) Acute Myeloid Leukemic Stem Cells by Restoring p53 Acetylation. Cell stem cell, 17(5), 597-610.

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Immunoprecipitation with Flag/M2 or Protein A/G beads

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Antibodies used for immunoprecipitation were conjugated with Flag/M2 beads (Sigma-Aldrich) or protein A/G beads using an antibody crosslinking kit (Pierce Biotechnology). Western blotting was performed as described in the Supplemental Experimental Procedures.

Li L., Osdal T., Ho Y., Chun S., McDonald T., Agarwal P., Lin A., Chu S., Qi J., Li L., Hsieh Y.T., Santos C.D., Yuan H., Ha T.Q., Popa M., Hovland R., Bruserud Ø., Gjertsen B.T., Kuo Y.H., Chen W., Lain S., McCormack E, & Bhatia R. (2014). SIRT1 Activation by a c-MYC Oncogenic Network Promotes the Maintenance and Drug Resistance of Human FLT3-ITD Acute Myeloid Leukemia Stem Cells. Cell stem cell, 15(4), 431-446.

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