Anti-Phospho-Mcl1 (Ser159+Thr163) Polyclonal Antibody is a Rabbit antibody targeting Phospho-Mcl1 (Ser159+Thr163). Anti-Phospho-Mcl1 (Ser159+Thr163) Polyclonal Antibody can be used in IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. 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-Mcl1 (Ser159+Thr163)) Polyclonal Antibody, Unconjugated (TMAB-10956) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
2. Blank control: K562.
Primary Antibody (green line): Rabbit Anti-Phospho-Mcl1 (Ser159+Thr163) antibody (TMAB-10956)
Dilution: 1 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℃. 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.
3. A549 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-Mcl1 (Ser159+Thr163)) polyclonal Antibody, Unconjugated (TMAB-10956) 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.

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

Polyclonal

KLH conjugated synthesised phosphopeptide: human Mcl 1 around the phosphorylation site of Ser159/Thr163

Human

MCL1

Induced myeloid leukemia cell differentiation protein Mcl-1

Involved in the regulation of apoptosis versus cell survival, and in the maintenance of viability but not of proliferation. Mediates its effects by interactions with a number of other regulators of apoptosis. Isoform 1 inhibits apoptosis. Isoform 2 promotes apoptosis.