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Mouse anti h3cit monoclonal antibody

Manufactured by BioCat

The Mouse anti-H3Cit monoclonal antibody is a laboratory reagent used for the detection and analysis of histone H3 proteins that have been citrullinated (converted from arginine to citrulline) at specific sites. This antibody is designed to specifically bind to the citrullinated form of histone H3, allowing researchers to study this post-translational modification and its biological implications.

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2 protocols using mouse anti h3cit monoclonal antibody

1

Quantification of Histone H3 Citrullination

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Human neutrophils were isolated as above and diluted to a density of 3.5×105/mL. Cells were incubated with compounds in 384-well micro-titre plates for 45 min at 37 °C, 5% CO2. Cells were then stimulated with 2 µM calcium ionophore (final concentration) for 60 min at 37 °C. The cells were fixed with 1.3% PFA for 45 min at RT, then permeabilised in PBS/2% Triton X-100 for 10 min. Cells were washed with PBST (PBS/0.1% Tween 20) and blocked in PBST/2% BSA for 16 h at 4 °C. H3 citrullination was measured using a mouse anti-H3Cit monoclonal antibody (BioCat, 133483, generated at GSK, 6 µg/mL) and a secondary Alexa Fluor 488 goat anti-mouse IgG (Life Technologies, A11001, 2 µg/mL) with parallel Hoechst 33342 staining (Life Technologies). Plates were imaged on an IN Cell Analyzer 2000 (GE Healthcare) and the FITC intensity in neutrophil nuclei was used as a measure of H3 citrullination as determined using the IN Cell Investigator software (GE Healthcare).
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2

Quantification of Histone H3 Citrullination

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human neutrophils were isolated as above and diluted to a density of 3.5×105/mL. Cells were incubated with compounds in 384-well micro-titre plates for 45 min at 37 °C, 5% CO2. Cells were then stimulated with 2 µM calcium ionophore (final concentration) for 60 min at 37 °C. The cells were fixed with 1.3% PFA for 45 min at RT, then permeabilised in PBS/2% Triton X-100 for 10 min. Cells were washed with PBST (PBS/0.1% Tween 20) and blocked in PBST/2% BSA for 16 h at 4 °C. H3 citrullination was measured using a mouse anti-H3Cit monoclonal antibody (BioCat, 133483, generated at GSK, 6 µg/mL) and a secondary Alexa Fluor 488 goat anti-mouse IgG (Life Technologies, A11001, 2 µg/mL) with parallel Hoechst 33342 staining (Life Technologies). Plates were imaged on an IN Cell Analyzer 2000 (GE Healthcare) and the FITC intensity in neutrophil nuclei was used as a measure of H3 citrullination as determined using the IN Cell Investigator software (GE Healthcare).
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