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Anti hdac3 antibody

Manufactured by Proteintech
Sourced in United States

The Anti-HDAC3 antibody is a specific antibody that recognizes and binds to the HDAC3 protein. HDAC3 is a member of the histone deacetylase family of enzymes, which are involved in the regulation of gene expression. This antibody can be used in various laboratory techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to detect and study the HDAC3 protein.

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2 protocols using anti hdac3 antibody

1

Western Blot Analysis of HDAC3

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Cell lysates were loaded in a 10% polyacrylamide gel for electrophoresis, followed by nitrocellulose membrane transference based on the standard Western blot assay protocol. 5% non-fat milk in the PBS for blocking was incubated at 37 °C for 1 h. A corresponding antibody against a specific protein (anti-HDAC3 antibody was obtained from Proteintech, USA) was added for incubation at 4 °C overnight and followed by the detection with an appropriate horseradish peroxidase conjugated secondary antibody (1/1000, Cell Signaling Technology, USA) at 37 °C for 1 h, as described in our previous study [12 (link)]. After the final PBS washing, signal was developed by ECL detection system and relative photographic density was quantitated by a gel documentation and analysis system (Alpha Imager 2000, Alpha Innotech Corporation, USA).
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2

Western Blot Analysis of Protein Acetylation

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Protein samples were separated by 8%–12% SDS-PAGE and transferred to nitrocellulose membranes (Millipore). After blocking with 5% milk in TBST for 1h, the membranes were incubated overnight at 4°C with primary antibodies. Subsequently, membranes were washed and incubated with secondary antibodies (1:10,000) and detected using the chemiluminescence method. The intensity of the bands was quantified by ImageJ software. Antibodies included anti-NLRP3 antibody (1:1,000, CST), anti-GSDMD antibody (1:1,000, Abcam), anti-N-GSDMD antibody (1:1,000, Abcam), anti-Capspase-1 p20 antibody (1:1,000, AdipoGen), anti-4-HNE antibody (1:1,000, Abcam), anti-HDAC3 antibody (1:1,000, Proteintech), anti-HADHA antibody (1:1,000, Abcam), anti-Ace-lys antibody (1:1,000, Abcam), anti-H3 antibody (1:1,000, Proteintech), anti-COX4 antibody (1:1,000, Abcam), anti-PCNA antibody (1:1,000, Abcam). Anti-GAPDH (1:5,000, Invitrogen) was used as an internal control.
For acetylation-immunoprecipitation, cells were lysed with immunoprecipitation buffer [supplemented with TSA (10 mM)] and sonicated. The samples were immunoprecipitated with protein A/G beads (Sigma) overnight at 4°C, washed three times in lysis buffer, resolved by loading buffer, and analyzed by Western blotting.
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