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Anti h3 clone d1h2

Manufactured by Cell Signaling Technology
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Anti-H3 (clone D1H2) is a monoclonal antibody that recognizes histone H3. It is designed for use in various applications, such as Western blotting, immunoprecipitation, and chromatin immunoprecipitation (ChIP).

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Lab products found in correlation

Enhanced chemiluminescence kit, by Bio-Rad (2 mentions) Anti ahr antibody, by Enzo Life Sciences (2 mentions) Anti β actin clone ac 15, by Merck Group (2 mentions)

2 protocols using anti h3 clone d1h2

1

Nuclear Fractionation and Western Blot Analysis

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For nuclear extraction, samples were prepared using the NE-PER nuclear cytoplasmic extraction kit (Pierce) according to the manufacturer’s instruction. For Western blot proteins were separated on 4–12% SDS-PAGE gel. Following electrophoresis, proteins were transferred to a PVDF membrane and subsequently blocked for 1h (5% nonfat dry milk in Tris-buffered saline containing 0.05% Tween 20) and then incubated with anti-AhR antibody (Enzo Life Sciences) overnight at 4oC. Blots were then washed 3 times in Tris-buffered saline containing 0.05% Tween20 and incubated for 1h with secondary antibody at room temperature 5 , 13 . All blots were stripped and reprobed with anti-β-actin (clone AC-15, Sigma) and anti-H3 (clone D1H2, Cell Signalling Technology) to ensure equal protein loading and purity of cytoplasmic and nuclear extracts. The protein bands were detected using an enhanced chemiluminescence kit (Bio-Rad).
Shinde R., Hezaveh K., Halaby M.J., Kloetgen A., Chakravarthy A., da Silva Medina T., Deol R., Manion K.P., Baglaenko Y., Eldh M., Lamorte S., Wallace D., Chodisetti S.B., Ravishankar B., Liu H., Chaudhary K., Munn D.H., Tsirigos A., Madaio M., Gabrielsson S., Touma Z., Wither J., De Carvalho D.D, & McGaha T.L. (2018). Apoptotic cell–induced, TLR9-dependent AhR activity is required for immunological tolerance and suppression of systemic lupus erythematosus in mice and humans. Nature immunology, 19(6), 571-582.
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2

Nuclear Fractionation and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
For nuclear extraction, samples were prepared using the NE-PER nuclear cytoplasmic extraction kit (Pierce) according to the manufacturer’s instruction. For Western blot proteins were separated on 4–12% SDS-PAGE gel. Following electrophoresis, proteins were transferred to a PVDF membrane and subsequently blocked for 1h (5% nonfat dry milk in Tris-buffered saline containing 0.05% Tween 20) and then incubated with anti-AhR antibody (Enzo Life Sciences) overnight at 4oC. Blots were then washed 3 times in Tris-buffered saline containing 0.05% Tween20 and incubated for 1h with secondary antibody at room temperature 5 , 13 . All blots were stripped and reprobed with anti-β-actin (clone AC-15, Sigma) and anti-H3 (clone D1H2, Cell Signalling Technology) to ensure equal protein loading and purity of cytoplasmic and nuclear extracts. The protein bands were detected using an enhanced chemiluminescence kit (Bio-Rad).
Shinde R., Hezaveh K., Halaby M.J., Kloetgen A., Chakravarthy A., da Silva Medina T., Deol R., Manion K.P., Baglaenko Y., Eldh M., Lamorte S., Wallace D., Chodisetti S.B., Ravishankar B., Liu H., Chaudhary K., Munn D.H., Tsirigos A., Madaio M., Gabrielsson S., Touma Z., Wither J., De Carvalho D.D, & McGaha T.L. (2018). Apoptotic cell–induced, TLR9-dependent AhR activity is required for immunological tolerance and suppression of systemic lupus erythematosus in mice and humans. Nature immunology, 19(6), 571-582.
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