Rabbit Anti-Phospho-MAP3K8 (Ser400) antibody

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MAP3K8 (phospho S400); P-MAP3K8/Tpl2 (Ser400); Phospho-MAP3K8(Ser400); Phospho-Tpl2 (Ser400); P-Tpl2 (Ser400);c COT; Cancer Osaka thyroid oncogene; Cancer Osaka thyroid oncogene; CCOT; COT; COT proto oncogene serine/threonine protein kinase; EST; ESTF; Ew

Cat:

SL3454R

Species Reactivity：

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

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human MAP3K8/Tpl2 around the phosphorylation site of Ser400:CQ(p-S)LD

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=1ug/TestIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: RAW264.7(Mouse) Cell Lysate at 30 ugPrimary: Anti-Phospho-MAP3K8 (Ser400) (SL3454R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 53 kDObserved band size: 53 kDSample: HepG2(Human) Cell Lysate at 30 ugPrimary: Anti-Phospho-MAP3K8 (Ser400) (SL3454R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 53 kDObserved band size: 53 kDSample: Spleen (Mouse) Lysate at 40 ugPrimary: Anti-Phospho-MAP3K8 (Ser400) (SL3454R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 53 kDObserved band size: 55 kD Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-MAP3K8 (Ser400)) Polyclonal Antibody, Unconjugated (SL3454R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Blank control: Hela. Primary Antibody (green line): Rabbit Anti-MAP3K8 antibody (SL3454R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min 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.

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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This gene is an oncogene that encodes a member of the serine/threonine protein kinase family. The encoded protein localizes to the cytoplasm and can activate both the MAP kinase and JNK kinase pathways. This protein was shown to activate IkappaB kinases, and thus induce the nuclear production of NF-kappaB. This protein was also found to promote the production of TNF-alpha and IL-2 during T lymphocyte activation. This gene may also utilize a downstream in-frame translation start codon, and thus produce an isoform containing a shorter N-terminus. The shorter isoform has been shown to display weaker transforming activity. Alternate splicing results in multiple transcript variants that encode the same protein. [provided by RefSeq, Sep 2011]

Function:
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.

Subunit:
Forms a ternary complex with NFKB1 and TNIP2.

Subcellular Location:
Cytoplasm.

Tissue Specificity:
Expressed in several normal tissues and human tumor-derived cell lines.

Post-translational modifications:
Autophosphorylated. Isoform 1 undergoes phosphorylation mainly on Ser residues, and isoform 2 on both Ser and Thr residues.

Similarity:
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
Contains 1 protein kinase domain.

SWISS:
P41279

Gene ID:
1326

Database links:

Entrez Gene: 1326 Human

Entrez Gene: 26410 Mouse

Entrez Gene: 116596 Rat

Omim: 191195 Human

SwissProt: P41279 Human

SwissProt: Q07174 Mouse

SwissProt: Q63562 Rat

Unigene: 432453 Human

Unigene: 3275 Mouse

Unigene: 9939 Rat

Picture

Sample:
HepG2(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-MAP3K8 (Ser400) (SL3454R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 53 kD

Sample:
Spleen (Mouse) Lysate at 40 ug
Primary: Anti-Phospho-MAP3K8 (Ser400) (SL3454R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 53 kD
Observed band size: 55 kD

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-MAP3K8 (Ser400)) Polyclonal Antibody, Unconjugated (SL3454R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-MAP3K8 antibody (SL3454R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min 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.

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