TCA-precipitated protein extracts were used to analyze Bim1-HA expression in wild type Bim1-HA or Bim1-HA mutants. For all figures, except Supplementary Figure 4c , overnight cultures of C. neoformans cells were diluted to an OD600 of 0.3 in 5 ml of fresh SC medium, left untreated or treated with the indicated concentration of BCS for the indicated times. For the data in Supplementary Figure 4c , overnight cultures of C. neoformans cells were diluted to an OD600 of 0.3 and grown to an OD600 of 3. After each time point TCA was added to a final concentration of 10%, cells pelleted, collected with 20% TCA and further processed as described. Supernatants from experiment in Supplementary Figure 4c were directly treated with loading buffer before SDS-PAGE and immunoblotting analysis. TCA protein extracts and supernatants were analyzed by immunoblotting with anti-HA (Y-11, polyclonal, Santa Cruz or SG-77, polyclonal, Invitrogen) and anti-H3 (D1H2, polyclonal, Cell Signaling) antibodies.
Anti h3 d1h2
Manufactured by Cell Signaling Technology
Anti-H3 (D1H2) is a monoclonal antibody that recognizes histone H3. It is designed for use in various applications such as western blotting, immunoprecipitation, and immunofluorescence.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using anti h3 d1h2
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TCA-based Bim1-HA protein extraction
2
TCA-based Bim1-HA protein extraction
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TCA-precipitated protein extracts were used to analyze Bim1-HA expression in wild type Bim1-HA or Bim1-HA mutants. For all figures, except Supplementary Figure 4c , overnight cultures of C. neoformans cells were diluted to an OD600 of 0.3 in 5 ml of fresh SC medium, left untreated or treated with the indicated concentration of BCS for the indicated times. For the data in Supplementary Figure 4c , overnight cultures of C. neoformans cells were diluted to an OD600 of 0.3 and grown to an OD600 of 3. After each time point TCA was added to a final concentration of 10%, cells pelleted, collected with 20% TCA and further processed as described. Supernatants from experiment in Supplementary Figure 4c were directly treated with loading buffer before SDS-PAGE and immunoblotting analysis. TCA protein extracts and supernatants were analyzed by immunoblotting with anti-HA (Y-11, polyclonal, Santa Cruz or SG-77, polyclonal, Invitrogen) and anti-H3 (D1H2, polyclonal, Cell Signaling) antibodies.
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