Anti-MCL1 Polyclonal Antibody is a Rabbit antibody targeting MCL1. Anti-MCL1 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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 (MCL1) Polyclonal Antibody, Unconjugated (TMAB-08663) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); 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 (MCL1) Polyclonal Antibody, Unconjugated (TMAB-08663) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Blank control: K562.
Primary Antibody (green line): Rabbit Anti-MCL1 antibody (TMAB-08663)
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.
4. Sample:
Lane 1: Lymph node (Mouse) Lysate at 40 μg
Lane 2: Spleen (Mouse) Lysate at 40 μg
Lane 3: Small intestine (Mouse) Lysate at 40 μg
Lane 4: NIH/3T3 (Mouse) Cell Lysate at 30 μg
Lane 5: Lymph node (Rat) Lysate at 40 μg
Lane 6: Spleen (Rat) Lysate at 40 μg
Lane 7: Small intestine (Rat) Lysate at 40 μg
Lane 8: Bone (Rat) Lysate at 40 μg
Lane 9: HL60 (Human) Cell Lysate at 30 μg
Lane 10: A431 (Human) Cell Lysate at 30 μg
Lane 11: K562 (Human) Cell Lysate at 30 μg
Lane 12: MDA-MB-231 (Human) Cell Lysate at 30 μg
Lane 13: Raji (Human) Cell Lysate at 30 μg
Lane 14: A549 (Human) Cell Lysate at 30 μg
Primary: Anti-MCL1 (TMAB-08663) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 39 kD
Observed band size: 40 kD
5. Paraformaldehyde-fixed, paraffin embedded (Mouse embryo); 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 (MCL1) Polyclonal Antibody, Unconjugated (TMAB-08663) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded (human lung carcinoma); 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 (MCL1) Polyclonal Antibody, Unconjugated (TMAB-08663) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

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

Polyclonal

KLH conjugated synthetic peptide: human MCL1

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.