Product: Phospho-PAK1 (Thr212) Antibody
Catalog: AF3424
Description: Rabbit polyclonal antibody to Phospho-PAK1 (Thr212)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 65kDa; 61kD(Calculated).
Uniprot: Q13153
RRID: AB_2834866

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*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(92%), Bovine(92%), Horse(92%), Sheep(92%), Rabbit(100%), Dog(92%)
Clonality:
Polyclonal
Specificity:
Phospho-PAK1 (Thr212) Antibody detects endogenous levels of PAK1 only when phosphorylated at Threonine 212.
RRID:
AB_2834866
Cite Format: Affinity Biosciences Cat# AF3424, RRID:AB_2834866.
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

ADRB2; Alpha PAK; Alpha-PAK; MGC130000; MGC130001; p21 activated kinase 1; p21 protein (Cdc42/Rac) activated kinase 1; p21-activated kinase 1; p21/Cdc42/Rac1 activated kinase 1 (yeast Ste20 related); p21/Cdc42/Rac1-activated kinase 1 (STE20 homolog, yeast); p65 PAK; p65-PAK; P68-PAK; PAK alpha; PAK-1; Pak1; PAK1_HUMAN; Paka; PAKalpha; Protein kinase MUK2; Rac/p21-activated kinase; Serine/threonine-protein kinase PAK 1; STE20 homolog yeast;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q13153 PAK1_HUMAN:

Overexpressed in gastric cancer cells and tissues (at protein level) (PubMed:25766321).

Description:
PAK proteins are critical effectors that link RhoGTPases to cytoskeleton reorganization and nuclear signaling. PAK proteins, a family of serine/threonine p21-activating kinases, include PAK1, PAK2, PAK3 and PAK4.
Sequence:
MSNNGLDIQDKPPAPPMRNTSTMIGAGSKDAGTLNHGSKPLPPNPEEKKKKDRFYRSILPGDKTNKKKEKERPEISLPSDFEHTIHVGFDAVTGEFTGMPEQWARLLQTSNITKSEQKKNPQAVLDVLEFYNSKKTSNSQKYMSFTDKSAEDYNSSNALNVKAVSETPAVPPVSEDEDDDDDDATPPPVIAPRPEHTKSVYTRSVIEPLPVTPTRDVATSPISPTENNTTPPDALTRNTEKQKKKPKMSDEEILEKLRSIVSVGDPKKKYTRFEKIGQGASGTVYTAMDVATGQEVAIKQMNLQQQPKKELIINEILVMRENKNPNIVNYLDSYLVGDELWVVMEYLAGGSLTDVVTETCMDEGQIAAVCRECLQALEFLHSNQVIHRDIKSDNILLGMDGSVKLTDFGFCAQITPEQSKRSTMVGTPYWMAPEVVTRKAYGPKVDIWSLGIMAIEMIEGEPPYLNENPLRALYLIATNGTPELQNPEKLSAIFRDFLNRCLEMDVEKRGSAKELLQHQFLKIAKPLSSLTPLIAAAKEATKNNH

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
Rabbit
100
Pig
92
Horse
92
Bovine
92
Sheep
92
Dog
92
Chicken
77
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13153 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
K11 Ubiquitination
T20 Phosphorylation
S21 Phosphorylation Q13153 (PAK1) , Q13976-2 (PRKG1) , P31749 (AKT1)
S28 Phosphorylation
K29 Acetylation
K29 Ubiquitination
S38 Phosphorylation
K39 Ubiquitination
S57 Phosphorylation Q13153 (PAK1)
K63 Acetylation
S79 Phosphorylation
T84 Phosphorylation O95747 (OXSR1)
T109 Phosphorylation Q15831 (STK11)
S110 Phosphorylation
S115 Phosphorylation
Y131 Phosphorylation
S133 Phosphorylation
T136 Phosphorylation
S137 Phosphorylation
S139 Phosphorylation
Y142 Phosphorylation
S144 Phosphorylation Q13153 (PAK1)
T146 Phosphorylation
K148 Ubiquitination
S149 Phosphorylation Q13153 (PAK1)
Y153 Phosphorylation O60674 (JAK2)
S155 Phosphorylation
S156 Phosphorylation
S174 Phosphorylation Q9BWU1 (CDK19)
T185 Phosphorylation
T197 Phosphorylation
S199 Phosphorylation Q13153 (PAK1)
Y201 Phosphorylation O60674 (JAK2)
S204 Phosphorylation Q13153 (PAK1)
T212 Phosphorylation P28482 (MAPK1)
T214 Phosphorylation
T219 Phosphorylation
S220 Phosphorylation
S223 Phosphorylation P68400 (CSNK2A1)
T225 Phosphorylation
T229 Phosphorylation
T230 Phosphorylation
T236 Phosphorylation
S249 Phosphorylation
K256 Acetylation
K256 Sumoylation
K256 Ubiquitination
S259 Phosphorylation
S262 Phosphorylation
Y285 Phosphorylation O60674 (JAK2)
T292 Phosphorylation P27361 (MAPK3)
K309 Ubiquitination
K391 Ubiquitination
T415 Phosphorylation
K420 Acetylation
K420 Ubiquitination
S422 Phosphorylation
T423 Phosphorylation O15530 (PDPK1) , Q8IWQ3 (BRSK2) , Q13153 (PAK1)
T427 Phosphorylation
Y474 Phosphorylation
S491 Phosphorylation
K508 Ubiquitination
S511 Phosphorylation
K513 Acetylation
S528 Phosphorylation
S529 Phosphorylation
T531 Phosphorylation

PTMs - Q13153 As Enzyme

Substrate Site Source
O14965 (AURKA) T288 Uniprot
O14965 (AURKA) S342 Uniprot
O15143 (ARPC1B) T21 Uniprot
O95863 (SNAI1) S246 Uniprot
P03372-1 (ESR1) S305 Uniprot
P04049 (RAF1) S338 Uniprot
P04049 (RAF1) S339 Uniprot
P08670 (VIM) S26 Uniprot
P08670 (VIM) S39 Uniprot
P08670 (VIM) S51 Uniprot
P08670 (VIM) S56 Uniprot
P08670 (VIM) S66 Uniprot
P08670 (VIM) S73 Uniprot
P10398 (ARAF) S299 Uniprot
P15056 (BRAF) S446 Uniprot
P16949 (STMN1) S16 Uniprot
P16949 (STMN1) S38 Uniprot
P17600-2 (SYN1) S605 Uniprot
P18669 (PGAM1) S23 Uniprot
P18669 (PGAM1) S118 Uniprot
P19086 (GNAZ) S16 Uniprot
P21333 (FLNA) S2152 Uniprot
P21333 (FLNA) S2292 Uniprot
P21333 (FLNA) S2370 Uniprot
P24844 (MYL9) S20 Uniprot
P29466-1 (CASP1) S376 Uniprot
P31749 (AKT1) S473 Uniprot
P35222 (CTNNB1) S675 Uniprot
P35240-4 (NF2) S435 Uniprot
P35240-6 (NF2) S476 Uniprot
P35240-5 (NF2) S477 Uniprot
P35240-3 (NF2) S518 Uniprot
P36871 (PGM1) T467 Uniprot
P42229 (STAT5A) S780 Uniprot
P46108 (CRK) S41 Uniprot
P49023 (PXN) S258 Uniprot
P49023-2 (PXN) S272 Uniprot
P51114 (FXR1) S420 Uniprot
P52565 (ARHGDIA) S101 Uniprot
P52565 (ARHGDIA) S174 Uniprot
P53350 (PLK1) S49 Uniprot
P53667 (LIMK1) T508 Uniprot
P58012 (FOXL2) S263 Uniprot
P63167 (DYNLL1) S88 Uniprot
P68431 (HIST1H3J) S11 Uniprot
P78545 (ELF3) S207 Uniprot
Q02750-1 (MAP2K1) S298 Uniprot
Q12778 (FOXO1) S256 Uniprot
Q13153-1 (PAK1) S21 Uniprot
Q13153-1 (PAK1) S57 Uniprot
Q13153 (PAK1) S144 Uniprot
Q13153 (PAK1) S149 Uniprot
Q13153-1 (PAK1) S165 Uniprot
Q13153 (PAK1) S199 Uniprot
Q13153-1 (PAK1) S204 Uniprot
Q13153 (PAK1) T423 Uniprot
Q13233 (MAP3K1) S67 Uniprot
Q13352 (ITGB3BP) S28 Uniprot
Q13363 (CTBP1) S158 Uniprot
Q13418 (ILK) T173 Uniprot
Q13418 (ILK) S246 Uniprot
Q14155 (ARHGEF7) S518 Uniprot
Q14247 (CTTN) S405 Uniprot
Q14247 (CTTN) S418 Uniprot
Q15365 (PCBP1) T60 Uniprot
Q15365 (PCBP1) T127 Uniprot
Q15746-1 (MYLK) S18 Uniprot
Q15746 (MYLK) S1772 Uniprot
Q70Z35 (PREX2) S1107 Uniprot
Q7Z628 (NET1) S152 Uniprot
Q8IYU2 (HACE1) S385 Uniprot
Q92934 (BAD) S74 Uniprot
Q92934 (BAD) S75 Uniprot
Q92934 (BAD) S99 Uniprot
Q92934 (BAD) S118 Uniprot
Q92934 (BAD) S134 Uniprot
Q92974-3 (ARHGEF2) S858 Uniprot
Q92974 (ARHGEF2) S886 Uniprot
Q96A00 (PPP1R14A) T38 Uniprot
Q96T58 (SPEN) S3486 Uniprot
Q96T58 (SPEN) T3568 Uniprot
Q99426 (TBCB) S65 Uniprot
Q99426 (TBCB) S128 Uniprot
Q99661 (KIF2C) S111 Uniprot
Q99661 (KIF2C) S192 Uniprot
Q9UQ80 (PA2G4) T261 Uniprot
Q9Y2X7 (GIT1) S508 Uniprot
Q9Y2X7 (GIT1) S700 Uniprot
Q9Y6X9 (MORC2) S739 Uniprot

Research Backgrounds

Function:

Protein kinase involved in intracellular signaling pathways downstream of integrins and receptor-type kinases that plays an important role in cytoskeleton dynamics, in cell adhesion, migration, proliferation, apoptosis, mitosis, and in vesicle-mediated transport processes. Can directly phosphorylate BAD and protects cells against apoptosis. Activated by interaction with CDC42 and RAC1. Functions as GTPase effector that links the Rho-related GTPases CDC42 and RAC1 to the JNK MAP kinase pathway. Phosphorylates and activates MAP2K1, and thereby mediates activation of downstream MAP kinases. Involved in the reorganization of the actin cytoskeleton, actin stress fibers and of focal adhesion complexes. Phosphorylates the tubulin chaperone TBCB and thereby plays a role in the regulation of microtubule biogenesis and organization of the tubulin cytoskeleton. Plays a role in the regulation of insulin secretion in response to elevated glucose levels. Part of a ternary complex that contains PAK1, DVL1 and MUSK that is important for MUSK-dependent regulation of AChR clustering during the formation of the neuromuscular junction (NMJ). Activity is inhibited in cells undergoing apoptosis, potentially due to binding of CDC2L1 and CDC2L2. Phosphorylates MYL9/MLC2. Phosphorylates RAF1 at 'Ser-338' and 'Ser-339' resulting in: activation of RAF1, stimulation of RAF1 translocation to mitochondria, phosphorylation of BAD by RAF1, and RAF1 binding to BCL2. Phosphorylates SNAI1 at 'Ser-246' promoting its transcriptional repressor activity by increasing its accumulation in the nucleus. In podocytes, promotes NR3C2 nuclear localization. Required for atypical chemokine receptor ACKR2-induced phosphorylation of LIMK1 and cofilin (CFL1) and for the up-regulation of ACKR2 from endosomal compartment to cell membrane, increasing its efficiency in chemokine uptake and degradation. In synapses, seems to mediate the regulation of F-actin cluster formation performed by SHANK3, maybe through CFL1 phosphorylation and inactivation. Plays a role in RUFY3-mediated facilitating gastric cancer cells migration and invasion. In response to DNA damage, phosphorylates MORC2 which activates its ATPase activity and facilitates chromatin remodeling.

PTMs:

Autophosphorylated in trans, meaning that in a dimer, one kinase molecule phosphorylates the other one. Activated by autophosphorylation at Thr-423 in response to a conformation change, triggered by interaction with GTP-bound CDC42 or RAC1. Activated by phosphorylation at Thr-423 by BRSK2 and by PDPK1. Phosphorylated by JAK2 in response to PRL; this increases PAK1 kinase activity. Phosphorylated at Ser-21 by PKB/AKT; this reduces interaction with NCK1 and association with focal adhesion sites. Upon DNA damage, phosphorylated at Thr-212 and translocates to the nucleoplasm. Phosphorylated at tyrosine residues, which can be enhanced by NTN1 (By similarity).

Subcellular Location:

Cytoplasm. Cell junction>Focal adhesion. Cell membrane. Cell projection>Ruffle membrane. Cell projection>Invadopodium. Nucleus>Nucleoplasm. Chromosome.
Note: Colocalizes with RUFY3, F-actin and other core migration components in invadopodia at the cell periphery (PubMed:25766321). Recruited to the cell membrane by interaction with CDC42 and RAC1. Recruited to focal adhesions upon activation. Colocalized with CIB1 within membrane ruffles during cell spreading upon readhesion to fibronectin. Upon DNA damage, translocates to the nucleoplasm when phosphorylated at Thr-212 where is co-recruited with MORC2 on damaged chromatin (PubMed:23260667).

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

Overexpressed in gastric cancer cells and tissues (at protein level).

Subunit Structure:

Homodimer; homodimerization results in autoinhibition. Active as monomer. Component of cytoplasmic complexes, which also contains PXN, ARHGEF6 and GIT1. Interacts with NISCH (By similarity). Interacts with DVL1; mediates the formation of a DVL1, MUSK and PAK1 ternary complex involved in AChR clustering (By similarity). Binds to the caspase-cleaved p110 isoform of CDC2L1 and CDC2L2, p110C, but not the full-length proteins. Interacts with ARHGEF7. Interacts tightly with GTP-bound but not GDP-bound CDC42/P21 and RAC1 (By similarity). Probably found in a ternary complex composed of DSCAM, PAK1 and RAC1. Interacts with DSCAM (via cytoplasmic domain); the interaction is direct and enhanced in presence of RAC1. Interacts with SCRIB. Interacts with PDPK1. Interacts (via kinase domain) with RAF1. Interacts with NCK1 and NCK2. Interacts with TBCB. Interacts with CRIPAK. Interacts with BRSK2 (By similarity). Interacts with SNAI1. Interacts with CIB1 isoform 2. Interacts with CIB1 (via N-terminal region); the interaction is direct, promotes PAK1 activity and occurs in a calcium-dependent manner. Interacts with INPP5K.

Family&Domains:

Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.   (View pathway)

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

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

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

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

· Environmental Information Processing > Signal transduction > Hippo signaling pathway - multiple species.   (View pathway)

· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Renal cell carcinoma.   (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Development > Axon guidance.   (View pathway)

· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.   (View pathway)

· Organismal Systems > Immune system > T cell receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.   (View pathway)

References

1). Calycosin inhibited autophagy and oxidative stress in chronic kidney disease skeletal muscle atrophy by regulating AMPK/SKP2/CARM1 signalling pathway. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, 2020 (PubMed: 32910538) [IF=5.3]

2). Salidroside ameliorates endothelial inflammation and oxidative stress by regulating the AMPK/NF-κB/NLRP3 signaling pathway in AGEs-induced HUVECs. European Journal of Pharmacology, 2020 (PubMed: 31747547) [IF=5.0]

Application: WB    Species: human    Sample: HUVECs

Fig. 8. |An AMPK inhibitor abrogated the protective effect of salidroside against inflammation in AGEs-induced HUVECs.(A) Representative western blot results for AMPK/NF-κB p65/NLRP3 signaling pathway proteins in AGEs/salidroside treated-HUVECs with or without compound C.GAPDH was used as a loading control. (B) Histograms analysis of the western blot results. The data are representative of one experiment performed in triplicate and are expressed as the mean ± S.D.

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