Anti-Phospho-HDAC5 (Ser498) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-HDAC5 (Ser498). Anti-Phospho-HDAC5 (Ser498) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse (predicted:Rat,Chicken,Dog,Pig,Cow,Horse,Rabbit)

1. Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (HDAC5 (Ser498)) Polyclonal Antibody, Unconjugated (TMAB-10774) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Sample: k562 (human) Cell Lysate at 30 μg
Primary: Anti-phospho-HDAC5 (Ser498) (TMAB-10774) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 123 kD
Observed band size: 123/112 kD
3. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-phospho-HDAC5 (Ser498) antibody (TMAB-10774)
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 HDAC5 around the phosphorylation site of Ser498

Human

HDAC5

Histone deacetylase 5

Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). 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. Involved in muscle maturation by repressing transcription of myocyte enhancer MEF2C. During muscle differentiation, it shuttles into the cytoplasm, allowing the expression of myocyte enhancer factors.