Anti-Phospho-MAP3K8 (Ser400) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-MAP3K8 (Ser400). Anti-Phospho-MAP3K8 (Ser400) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Sample: Spleen (Mouse) Lysate at 40 μg
Primary: Anti-Phospho-MAP3 k8 (Ser400) (TMAB-10932) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 55 kD
2. Paraformaldehyde-fixed, paraffin embedded (Rat 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-MAP3 k8 (Ser400)) Polyclonal Antibody, Unconjugated (TMAB-10932) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-MAP3 k8 antibody (TMAB-10932)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 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 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: RAW264.7 (Mouse) Cell Lysate at 30 μg
Primary: Anti-Phospho-MAP3 k8 (Ser400) (TMAB-10932) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 53 kD
5. Sample: HepG2 (Human) Cell Lysate at 30 μg
Primary: Anti-Phospho-MAP3 k8 (Ser400) (TMAB-10932) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 53 kD

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

Polyclonal

KLH conjugated Synthesised phosphopeptide: human MAP3K8/Tpl2 around the phosphorylation site of Ser400

Human

MAP3K8

Mitogen-activated protein kinase kinase kinase 8

Required for TLR4 activation of the MEK/ERK pathway. Able to activate NF-kappa-B 1 by stimulating proteasome-mediated proteolysis of NF-kappa-B 1/p105. Plays a role in the cell cycle. The longer form has some transforming activity, although it is much weaker than the activated cot oncoprotein.