Anti-Phospho-HDAC3 (Ser424) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-HDAC3 (Ser424). Anti-Phospho-HDAC3 (Ser424) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. Tissue/cell: Rat rectum tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Phospho-HDAC3, Unconjugated (TMAB-10769) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Sample:
MCF-7 (Human) Cell Lysate at 30 μg
NIH/3T3 (Mouse) Cell Lysate at 30 μg
Hela (Human) Cell Lysate at 30 μg
Primary: Anti-Phospho-HDAC3 (Ser424) (TMAB-10769) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD
3. Blank control (black line): Molt4.
Primary Antibody (green line): Rabbit Anti-Phospho-HDAC3 (Ser424) antibody (TMAB-10769)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
4. Sample:
Lane 1: Mouse NIH/3T3 cell lysates
Lane 2: Mouse Testis tissue lysates
Lane 3: Mouse Cerebrum tissue lysates
Lane 4: Rat Cerebrum tissue lysates
Primary: Anti-Phospho-HDAC3 (Ser424) (TMAB-10769) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 47 kDa
Observed band size: 50 kDa
5. Blank control (black line): NIH/3T3.
Primary Antibody (green line): Rabbit Anti-Phospho-HDAC3 (Ser424) antibody (TMAB-10769)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human HDAC3 around the phosphorylation site of Ser424

Human

HDAC3

Histone deacetylase 3

Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4), and some other non-histone substrates. Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Probably participates in the regulation of transcription through its binding to the zinc-finger transcription factor YY1; increases YY1 repression activity. Required to repress transcription of the POU1F1 transcription factor. Acts as a molecular chaperone for shuttling phosphorylated NR2C1 to PML bodies for sumoylation.