Anti-Phospho-MLK3 (Thr277+Ser281) Polyclonal Antibody is a Rabbit antibody targeting Phospho-MLK3 (Thr277+Ser281). Anti-Phospho-MLK3 (Thr277+Ser281) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Cow,Rabbit,GuineaPig)

1. Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); 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 (M3 k11) Polyclonal Antibody, Unconjugated (TMAB-10999) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
2. 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 (Phospho-MLK3 (Thr277+Ser281)) Polyclonal Antibody, Unconjugated (TMAB-10999) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (rat liver); 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-MLK3 (Thr277+Ser281)) Polyclonal Antibody, Unconjugated (TMAB-10999) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
4. Blank control: A431.
Primary Antibody (green line): Rabbit Anti-MLK3 (Thr277+Ser281) antibody (TMAB-10999)
Dilution: 3 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 3 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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.
5. Blank control (Black line): A431 (Black).
Primary Antibody (green line): Rabbit Anti-MLK3 antibody (TMAB-10999)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): 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 0.1% PBST for 20 min at room temperature. 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.

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

Polyclonal

KLH conjugated synthesised phosphopeptide: human MLK3 around the phosphorylation site of Thr277/Ser281

Human

MAP3K11

Mitogen-activated protein kinase kinase kinase 11

Activates the JUN N-terminal pathway. Required for serum-stimulated cell proliferation and for mitogen and cytokine activation of MAPK14 (p38), MAPK3 (ERK) and MAPK8 (JNK1). Plays a role in mitogen-stimulated phosphorylation and activation of BRAF, but does not phosphorylate BRAF directly. Influences microtubule organization during the cell cycle.