Mst1(Thr183) Polyclonal Antibody

L1 rat brain lysates L2 rat kidney lysates probed with Anti Phospho-Mst1(Thr183) Polyclonal Antibody, Unconjugated (bs-4635R) at 1:200 overnight at 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 for 90 min at RT. Predicted band 56kD. Observed band size:56kD.\\n

Antigen: bs-4635P, 0.2ug\/100ul \\nPrimary: Antiserum, 1:500, 1:1000, 1:2000, 1:4000, 1:8000, 1:16000, 1:32000; \\nSecondary: HRP conjugated Goat-Anti-Rabbit IgG(bs-0295G-HRP) at 1: 5000;\\nTMB staining;\\nRead the data in MicroplateReader by 450

HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Phospho-MST1 (Thr183)) polyclonal Antibody, Unconjugated (bs-4635R) 1:25, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

HepG2 cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with Phospho-MST1 (Thr183) Antibody(bs-4635R)at 1:50 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).