Phospho-MLK3 (Thr277+Ser281) Antibody - #AF8079

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Product: Phospho-MLK3 (Thr277+Ser281) Antibody
Catalog: AF8079
Description: Rabbit polyclonal antibody to Phospho-MLK3 (Thr277+Ser281)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 92KD; 93kD(Calculated).
Uniprot: Q16584
RRID: AB_2840142

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 100ul $350 In stock
 200ul $450 In stock

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-MLK3 (Thr277+Ser281) Antibody detects endogenous levels of MLK3 only when phosphorylated at Thr277+Ser281.
RRID:
AB_2840142
Cite Format: Affinity Biosciences Cat# AF8079, RRID:AB_2840142.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

2610017K16Rik; EC 2.7.11.25; M3K11_HUMAN; Map3k11; MEKK11; MGC17114; Mitogen-activated protein kinase kinase kinase 11; Mixed lineage kinase 3; Mixed lineage protein kinase 3; Mlk-3; Mlk3; Protein tyrosine kinase PTK1; PTK1; RHOE; SH3 domain containing proline rich kinase; SPRK; Src-homology 3 domain-containing proline-rich kinase;

Immunogens

Immunogen:
Uniprot:

Q16584(M3K11_HUMAN) >>Visit HPA database.

Gene(ID):
MAP3K11(4296)
Expression:
Q16584 M3K11_HUMAN:

Expressed in a wide variety of normal and neoplastic tissues including fetal lung, liver, heart and kidney, and adult lung, liver, heart, kidney, placenta, skeletal muscle, pancreas and brain.

Sequence:
MEPLKSLFLKSPLGSWNGSGSGGGGGGGGGRPEGSPKAAGYANPVWTALFDYEPSGQDELALRKGDRVEVLSRDAAISGDEGWWAGQVGGQVGIFPSNYVSRGGGPPPCEVASFQELRLEEVIGIGGFGKVYRGSWRGELVAVKAARQDPDEDISVTAESVRQEARLFAMLAHPNIIALKAVCLEEPNLCLVMEYAAGGPLSRALAGRRVPPHVLVNWAVQIARGMHYLHCEALVPVIHRDLKSNNILLLQPIESDDMEHKTLKITDFGLAREWHKTTQMSAAGTYAWMAPEVIKASTFSKGSDVWSFGVLLWELLTGEVPYRGIDCLAVAYGVAVNKLTLPIPSTCPEPFAQLMADCWAQDPHRRPDFASILQQLEALEAQVLREMPRDSFHSMQEGWKREIQGLFDELRAKEKELLSREEELTRAAREQRSQAEQLRRREHLLAQWELEVFERELTLLLQQVDRERPHVRRRRGTFKRSKLRARDGGERISMPLDFKHRITVQASPGLDRRRNVFEVGPGDSPTFPRFRAIQLEPAEPGQAWGRQSPRRLEDSSNGERRACWAWGPSSPKPGEAQNGRRRSRMDEATWYLDSDDSSPLGSPSTPPALNGNPPRPSLEPEEPKRPVPAERGSSSGTPKLIQRALLRGTALLASLGLGRDLQPPGGPGRERGESPTTPPTPTPAPCPTEPPPSPLICFSLKTPDSPPTPAPLLLDLGIPVGQRSAKSPRREEEPRGGTVSPPPGTSRSAPGTPGTPRSPPLGLISRPRPSPLRSRIDPWSFVSAGPRPSPLPSPQPAPRRAPWTLFPDSDPFWDSPPANPFQGGPQDCRAQTKDMGAQAPWVPEAGP

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16584 As Substrate

Site PTM Type Enzyme
S11 Phosphorylation
S35 Phosphorylation
K261 Ubiquitination
K264 Ubiquitination
T277 Phosphorylation Q16584 (MAP3K11)
S281 Phosphorylation Q16584 (MAP3K11) , Q92918 (MAP4K1)
S394 Phosphorylation
T477 Phosphorylation
S493 Phosphorylation
K499 Ubiquitination
S507 Phosphorylation
R512 Methylation
S524 Phosphorylation
T526 Phosphorylation
S548 Phosphorylation
S555 Phosphorylation
S556 Phosphorylation
S570 Phosphorylation
K624 Ubiquitination
S654 Phosphorylation
S674 Phosphorylation P31749 (AKT1)
T677 Phosphorylation
S693 Phosphorylation
T702 Phosphorylation
S705 Phosphorylation
T708 Phosphorylation
S724 Phosphorylation
S727 Phosphorylation
T738 Phosphorylation
S740 Phosphorylation
S746 Phosphorylation
S748 Phosphorylation
T752 Phosphorylation
T755 Phosphorylation
S758 Phosphorylation
S770 Phosphorylation
S774 Phosphorylation
S783 Phosphorylation
S789 Phosphorylation P49840 (GSK3A)
S793 Phosphorylation P49840 (GSK3A)
S815 Phosphorylation

PTMs - Q16584 As Enzyme

Substrate Site Source
P45985 (MAP2K4) S257 Uniprot
P45985 (MAP2K4) T261 Uniprot
P46734 (MAP2K3) S218 Uniprot
P46734 (MAP2K3) T222 Uniprot
Q13131 (PRKAA1) T183 Uniprot
Q13526 (PIN1) S138 Uniprot
Q16584 (MAP3K11) T277 Uniprot
Q16584 (MAP3K11) S281 Uniprot

Research Backgrounds

Function:

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) through phosphorylation and activation of MAP2K4/MKK4 and MAP2K7/MKK7. Plays a role in mitogen-stimulated phosphorylation and activation of BRAF, but does not phosphorylate BRAF directly. Influences microtubule organization during the cell cycle.

PTMs:

Autophosphorylation on serine and threonine residues within the activation loop plays a role in enzyme activation. Thr-277 is likely to be the main autophosphorylation site. Phosphorylation of Ser-555 and Ser-556 is induced by CDC42.

Subcellular Location:

Cytoplasm>Cytoskeleton>Microtubule organizing center>Centrosome.
Note: Location is cell cycle dependent.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in a wide variety of normal and neoplastic tissues including fetal lung, liver, heart and kidney, and adult lung, liver, heart, kidney, placenta, skeletal muscle, pancreas and brain.

Subunit Structure:

Homodimer; undergoes dimerization during activation. Interacts with MAP2K4/MKK4 (By similarity). Interacts with MAP2K7/MKK7. Found in a complex with SH3RF1, RAC1, MAP2K7/MKK7, MAPK8IP1/JIP1 and MAPK8/JNK1 (By similarity).

Family&Domains:

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.

Research Fields

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

References

1). Targeting GPR65 alleviates hepatic inflammation and fibrosis by suppressing the JNK and NF-κB pathways. Military Medical Research, 2023 (PubMed: 38001521) [IF=21.1]

Application: WB    Species: Mouse    Sample: liver tissues

Fig. 6 Targeting GPR65 alleviates hepatic macrophage inflammation and fibrosis by suppressing the JNK/NF-κB pathways. a Western blotting was used to determine the level of p-IKK, p-IĸBα, p-NF-κB p65, IKK, IĸBα, NF-κB p65, p-MLK3, p-MKK4, p-MKK7, p-JNK, JNK, p-p44/42, p44/42, p-p38, p38, p-GSK-3β, p-Akt, Akt, p-PDK1, p-PTEN and p–c-Raf in liver tissues from WT, WT + BDL, GPR65-KO and GPR65-KO + BDL mice. b Western blotting was used to determine the level of p-IKK, IKK, p-IĸBα, IĸBα, p-p65, p65, p-MLK3, p-MKK7, p-JNK and JNK in GPR65-KO, GPR65-overexpressed, various pH-treated and GPR65 agonist/inhibitor-treated HMs. The specific inhibitors of JNK, SP600125 (10 μmol/L) and JNK-IN-8 (5 μmol/L), as well as the specific inhibitors of NF-κB, BAY 11–7082 (5 μmol/L) and APDC (20 μmol/L), were used to treat GPR65-overexpressed HMs for 24 h, and qRT-PCR was used to assess the expression of Tnfα, Il6 and Tgfβ3 (n = 3) (c); TGF-β1 and IL-6 level in the supernatant were detected by ELISA (n = 3) (d); the expression and location of TNF-α was assessed by confocal microscopy (e). Scale bar = 20 μm. *P 
2). Disrupting PIAS3-mediated SUMOylation of MLK3 ameliorates poststroke neuronal damage and deficits in cognitive and sensorimotor behaviors. Cellular and molecular life sciences : CMLS, 2024 (PubMed: 38456949) [IF=8.0]

Application: WB    Species: Rat    Sample:

Fig. 3 Overexpressing the PINIT domain of PIAS3 blocks the MLK3-PIAS3 interaction and inhibits MLK3 SUMOylation and downstream MLK3-JNK/p38 apoptotic cascade activation after OGD/R. A Schematic diagram of blocking PIAS3-mediated MLK3 SUMOylation by the PINIT domain in the OGD model. B Diagram of lentiviral plasmids expressing the PINIT domain and negative control. C Lentivirus infection efficiency (green) in primary cortical neurons. Bar = 50 μm. D Schematic diagram of the overexpression of the PINIT domain in the primary neuron OGD model. E–G The effects of overexpressing the PINIT domain on the MLK3-PIAS3 interaction, MLK3 SUMOylation (E), MLK3 activation/phosphorylation (F), and downstream JNK/p38 apoptotic signaling (G) after OGD/R. − , without lentivirus infection. Relative levels were normalized to the normoxia group. Data are presented as the mean ± SD (n = 3–4). *P 

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