Anti-Phospho-MYPT1 (Thr696) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-MYPT1 (Thr696). Anti-Phospho-MYPT1 (Thr696) Polyclonal Antibody 2 can be used in ICC/IF,FCM.

IgG

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

1. Blank control (black line): HepG2.
Primary Antibody (green line): Rabbit Anti-Phospho-MYPT1 (Thr696) antibody (TMAB-11023)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Negative control (white blue line): PBS
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.
2. HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-MYPT1 (Thr696)) polyclonal Antibody, Unconjugated (TMAB-11023) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

ICC/IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human MYPT1 around the phosphorylation site of Thr696

Human

PPP1R12A

Protein phosphatase 1 regulatory subunit 12A

Key regulator of protein phosphatase 1C (PPP1C). Mediates binding to myosin. As part of the PPP1C complex, involved in dephosphorylation of PLK1. Capable of inhibiting HIF1AN-dependent suppression of HIF1A activity.