Anti-Phospho-Histone H1.4 (Thr18) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Histone H1.4 (Thr18). Anti-Phospho-Histone H1.4 (Thr18) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. Tissue/cell: rat brain 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-Histone H1.4 (Thr18) Polyclonal Antibody, Unconjugated (TMAB-10791) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Paraformaldehyde-fixed, paraffin embedded (rat 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 H1.4 (Thr18)) Polyclonal Antibody, Unconjugated (TMAB-10791) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (rat liver); 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 H1.4 (Thr18)) Polyclonal Antibody, Unconjugated (TMAB-10791) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 H1.4 (Thr18)) Polyclonal Antibody, Unconjugated (TMAB-10791) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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 H1.4 (Thr18)) Polyclonal Antibody, Unconjugated (TMAB-10791) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-phospho-Histone H1.4 (Thr18) antibody (TMAB-10791)
Dilution: 2 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Histone H1.4 around the phosphorylation site of Thr18

Human

H1-4

Histone H1.4

Histones H1 are necessary for the condensation of nucleosome chains into higher order structures.