Anti-Phospho-Histone H3 (Ser10) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-Histone H3 (Ser10). Anti-Phospho-Histone H3 (Ser10) Polyclonal Antibody 2 can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Rat (predicted:Mouse,Pig,Cow,Rabbit,Fruit Fly)

1. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-Phospho-Histone H3 (Ser10) antibody (TMAB-10796), Dilution: 0.2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg /test.
2. Paraformaldehyde-fixed, paraffin embedded (Human breast carcinoma); 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 (Phospho-Histone H3 (Ser10)) Polyclonal Antibody, Unconjugated (TMAB-10796) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-Histone H3 (Di Methyl K36) antibody (TMAB-10796-FITC)
Dilution: 2 μg /10^6 cells;
Isotype Control Antibody (orange line): 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. Acquisition of 20,000 events was performed.
4. Paraformaldehyde-fixed, paraffin embedded (Human kidney); 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 (Phospho-Histone H3 (Ser10)) Polyclonal Antibody, Unconjugated (TMAB-10796) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (Rat colon); 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 (Phospho-Histone H3 (Ser10)) Polyclonal Antibody, Unconjugated (TMAB-10796) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Blank control: U-87MG.
Primary Antibody (green line): Rabbit Anti-Phospho-Histone H3 antibody (TMAB-10796)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg /test.
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 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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 0.2μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human Histone H3 around the phosphorylation site of Ser10

Human

H3C1

Histone H3.1

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. H3 is deposited into chromatin exclusively through a DNA replication-coupled pathway that can be associated with either DNA duplication or DNA repair synthesis during meiotic homologous recombination.