Antibody Type: Rabbit Polyclonal

Application: WB,FCM

Reactivity: Human

IgG

Human

1. Blank control (blue line): HL60 cells (blue).
Primary Antibody (green line): Rabbit Anti-PARP antibody (TMAB-01335)
Dilution: 0.2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. Blank control: 293T.
Primary Antibody (green line): Rabbit Anti-PARP1 antibody (TMAB-01335)
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°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. Sample:
Lane 1: 293T (Human) Cell Lysate at 30 μg
Lane 2: HL60 (Human) Cell Lysate at 30 μg
Lane 3: K562 (Human) Cell Lysate at 30 μg
Lane 4: SH-SY5Y (Human) Cell Lysate at 30 μg
Lane 5: U251 (Human) Cell Lysate at 30 μg
Lane 6: Hela (Human) Cell Lysate at 30 μg
Primary: Anti-PARP (TMAB-01335) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 115 kDa
Observed band size: 115 kDa

WB,FCM

WB: 1:500-2000; FCM: 0.2μg/Test

Polyclonal

KLH conjugated synthetic peptide: human PARP

Human

142

ADP-ribosylation,Base Excision Repair,DNA Damage Recognition