Anti-Phospho-H2AFX (Ser139) Polyclonal Antibody is a Rabbit antibody targeting Phospho-H2AFX (Ser139). Anti-Phospho-H2AFX (Ser139) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Tissue/cell: human bladder carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Phospho-Histone H2A.x (Ser139) Polyclonal Antibody, Unconjugated (TMAB-01418) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Blank control: Molt4. Primary Antibody (green line): Rabbit Anti-Phospho-Histone H2A.X (Ser139) antibody
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-APC
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°C. 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.
3. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-Phospho-Histone H2A.X (Ser139) antibody (TMAB-01418)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
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°C. 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.
4. Sample:
Lane 1: Mouse BV2 cell lysates
Lane 2: Rat Testis tissue lysates
Lane 3: Human 293T cell lysates
Lane 4: Human Jurkat cell lysates
Primary: Anti-Phospho-H2AX (Ser139) (TMAB-01418) at 1/5000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 16 kDa
Observed band size: 15 kDa

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2ug/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human Histone H2AX around the phosphorylation site of Tyr139

Human

ChIP antibodies,DNA Damage Recognition,Unmodified,Variants