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$450 200ul

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  • Product Name
    Phospho-MLCK (Tyr464) Antibody
  • Catalog No.
    AF8387
  • RRID
    AB_2840448
  • Source
    Rabbit
  • Application
    WB,ELISA
  • Reactivity
    Human
  • UniProt
  • Mol.Wt
    211kD.;
    211kDa(Calculated).
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

deglutamylated form; DKFZp686I10125; EC 2.7.11.18; FLJ12216; Kinase related protein; Kinase-related protein; KRP; MLCK; MLCK1; MLCK108; MLCK210; MSTP083; MYLK; MYLK_HUMAN; MYLK1; Myosin light chain kinase; Myosin light polypeptide kinase; OTTHUMP00000180642; OTTHUMP00000180643; smMLCK; smooth muscle; Smooth muscle myosin light chain kinase; Telokin;

Applications:

WB 1:1000-3000, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.

Reactivity:

Human

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-MLCK (Tyr464) Antibody detects endogenous levels of MLCK only when phosphorylated at Tyr464.

RRID:

AB_2840448
Please cite this product as: Affinity Biosciences Cat# AF8387, RRID:AB_2840448.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

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.

Immunogen Information

Immunogen:

A synthesized peptide derived from human MLCK around the phosphorylation site of Tyr464.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

MYLK

Molecular Weight:

Observed Mol.Wt.: 211kD..
Predicted Mol.Wt.: 211kDa(Calculated)..

Tissue Specificity:

Q15746 MYLK_HUMAN:
Smooth muscle and non-muscle isozymes are expressed in a wide variety of adult and fetal tissues and in cultured endothelium with qualitative expression appearing to be neither tissue- nor development-specific. Non-muscle isoform 2 is the dominant splice variant expressed in various tissues. Telokin has been found in a wide variety of adult and fetal tissues. Accumulates in well differentiated enterocytes of the intestinal epithelium in response to tumor necrosis factor (TNF).

Sequence:
MGDVKLVASSHISKTSLSVDPSRVDSMPLTEAPAFILPPRNLCIKEGATAKFEGRVRGYPEPQVTWHRNGQPITSGGRFLLDCGIRGTFSLVIHAVHEEDRGKYTCEATNGSGARQVTVELTVEGSFAKQLGQPVVSKTLGDRFSAPAVETRPSIWGECPPKFATKLGRVVVKEGQMGRFSCKITGRPQPQVTWLKGNVPLQPSARVSVSEKNGMQVLEIHGVNQDDVGVYTCLVVNGSGKASMSAELSIQGLDSANRSFVRETKATNSDVRKEVTNVISKESKLDSLEAAAKSKNCSSPQRGGSPPWAANSQPQPPRESKLESCKDSPRTAPQTPVLQKTSSSITLQAARVQPEPRAPGLGVLSPSGEERKRPAPPRPATFPTRQPGLGSQDVVSKAANRRIPMEGQRDSAFPKFESKPQSQEVKENQTVKFRCEVSGIPKPEVAWFLEGTPVRRQEGSIEVYEDAGSHYLCLLKARTRDSGTYSCTASNAQGQLSCSWTLQVERLAVMEVAPSFSSVLKDCAVIEGQDFVLQCSVRGTPVPRITWLLNGQPIQYARSTCEAGVAELHIQDALPEDHGTYTCLAENALGQVSCSAWVTVHEKKSSRKSEYLLPVAPSKPTAPIFLQGLSDLKVMDGSQVTMTVQVSGNPPPEVIWLHNGNEIQESEDFHFEQRGTQHSLCIQEVFPEDTGTYTCEAWNSAGEVRTQAVLTVQEPHDGTQPWFISKPRSVTASLGQSVLISCAIAGDPFPTVHWLRDGKALCKDTGHFEVLQNEDVFTLVLKKVQPWHAGQYEILLKNRVGECSCQVSLMLQNSSARALPRGREPASCEDLCGGGVGADGGGSDRYGSLRPGWPARGQGWLEEEDGEDVRGVLKRRVETRQHTEEAIRQQEVEQLDFRDLLGKKVSTKTLSEDDLKEIPAEQMDFRANLQRQVKPKTVSEEERKVHSPQQVDFRSVLAKKGTSKTPVPEKVPPPKPATPDFRSVLGGKKKLPAENGSSSAETLNAKAVESSKPLSNAQPSGPLKPVGNAKPAETLKPMGNAKPAETLKPMGNAKPDENLKSASKEELKKDVKNDVNCKRGHAGTTDNEKRSESQGTAPAFKQKLQDVHVAEGKKLLLQCQVSSDPPATIIWTLNGKTLKTTKFIILSQEGSLCSVSIEKALPEDRGLYKCVAKNDAGQAECSCQVTVDDAPASENTKAPEMKSRRPKSSLPPVLGTESDATVKKKPAPKTPPKAAMPPQIIQFPEDQKVRAGESVELFGKVTGTQPITCTWMKFRKQIQESEHMKVENSENGSKLTILAARQEHCGCYTLLVENKLGSRQAQVNLTVVDKPDPPAGTPCASDIRSSSLTLSWYGSSYDGGSAVQSYSIEIWDSANKTWKELATCRSTSFNVQDLLPDHEYKFRVRAINVYGTSEPSQESELTTVGEKPEEPKDEVEVSDDDEKEPEVDYRTVTINTEQKVSDFYDIEERLGSGKFGQVFRLVEKKTRKVWAGKFFKAYSAKEKENIRQEISIMNCLHHPKLVQCVDAFEEKANIVMVLEIVSGGELFERIIDEDFELTERECIKYMRQISEGVEYIHKQGIVHLDLKPENIMCVNKTGTRIKLIDFGLARRLENAGSLKVLFGTPEFVAPEVINYEPIGYATDMWSIGVICYILVSGLSPFMGDNDNETLANVTSATWDFDDEAFDEISDDAKDFISNLLKKDMKNRLDCTQCLQHPWLMKDTKNMEAKKLSKDRMKKYMARRKWQKTGNAVRAIGRLSSMAMISGLSGRKSSTGSPTSPLNAEKLESEEDVSQAFLEAVAEEKPHVKPYFSKTIRDLEVVEGSAARFDCKIEGYPDPEVVWFKDDQSIRESRHFQIDYDEDGNCSLIISDVCGDDDAKYTCKAVNSLGEATCTAELIVETMEEGEGEGEEEEE

Research Background

Function:

Calcium/calmodulin-dependent myosin light chain kinase implicated in smooth muscle contraction via phosphorylation of myosin light chains (MLC). Also regulates actin-myosin interaction through a non-kinase activity. Phosphorylates PTK2B/PYK2 and myosin light-chains. Involved in the inflammatory response (e.g. apoptosis, vascular permeability, leukocyte diapedesis), cell motility and morphology, airway hyperreactivity and other activities relevant to asthma. Required for tonic airway smooth muscle contraction that is necessary for physiological and asthmatic airway resistance. Necessary for gastrointestinal motility. Implicated in the regulation of endothelial as well as vascular permeability, probably via the regulation of cytoskeletal rearrangements. In the nervous system it has been shown to control the growth initiation of astrocytic processes in culture and to participate in transmitter release at synapses formed between cultured sympathetic ganglion cells. Critical participant in signaling sequences that result in fibroblast apoptosis. Plays a role in the regulation of epithelial cell survival. Required for epithelial wound healing, especially during actomyosin ring contraction during purse-string wound closure. Mediates RhoA-dependent membrane blebbing. Triggers TRPC5 channel activity in a calcium-dependent signaling, by inducing its subcellular localization at the plasma membrane. Promotes cell migration (including tumor cells) and tumor metastasis. PTK2B/PYK2 activation by phosphorylation mediates ITGB2 activation and is thus essential to trigger neutrophil transmigration during acute lung injury (ALI). May regulate optic nerve head astrocyte migration. Probably involved in mitotic cytoskeletal regulation. Regulates tight junction probably by modulating ZO-1 exchange in the perijunctional actomyosin ring. Mediates burn-induced microvascular barrier injury; triggers endothelial contraction in the development of microvascular hyperpermeability by phosphorylating MLC. Essential for intestinal barrier dysfunction. Mediates Giardia spp.-mediated reduced epithelial barrier function during giardiasis intestinal infection via reorganization of cytoskeletal F-actin and tight junctional ZO-1. Necessary for hypotonicity-induced Ca(2+) entry and subsequent activation of volume-sensitive organic osmolyte/anion channels (VSOAC) in cervical cancer cells. Responsible for high proliferative ability of breast cancer cells through anti-apoptosis.

Post-translational Modifications:

Can probably be down-regulated by phosphorylation. Tyrosine phosphorylation by ABL1 increases kinase activity, reverses MLCK-mediated inhibition of Arp2/3-mediated actin polymerization, and enhances CTTN-binding. Phosphorylation by SRC at Tyr-464 and Tyr-471 promotes CTTN binding.

The C-terminus is deglutamylated by AGTPBP1/CCP1, AGBL1/CCP4 and AGBL4/CCP6, leading to the formation of Myosin light chain kinase, smooth muscle, deglutamylated form. The consequences of C-terminal deglutamylation are unknown (By similarity).

Acetylated at Lys-608 by NAA10/ARD1 via a calcium-dependent signaling; this acetylation represses kinase activity and reduces tumor cell migration.

Subcellular Location:

Cytoplasm. Cell projection>Lamellipodium. Cleavage furrow. Cytoplasm>Cytoskeleton>Stress fiber.
Note: Localized to stress fibers during interphase and to the cleavage furrow during mitosis.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Tissue Specificity:

Smooth muscle and non-muscle isozymes are expressed in a wide variety of adult and fetal tissues and in cultured endothelium with qualitative expression appearing to be neither tissue- nor development-specific. Non-muscle isoform 2 is the dominant splice variant expressed in various tissues. Telokin has been found in a wide variety of adult and fetal tissues. Accumulates in well differentiated enterocytes of the intestinal epithelium in response to tumor necrosis factor (TNF).

Subunit Structure:

All isoforms including Telokin bind calmodulin. Interacts with SVIL (By similarity). Interacts with CTTN; this interaction is reduced during thrombin-induced endothelial cell (EC) contraction but is promoted by the barrier-protective agonist sphingosine 1-phosphate (S1P) within lamellipodia. A complex made of ABL1, CTTN and MYLK regulates cortical actin-based cytoskeletal rearrangement critical to sphingosine 1-phosphate (S1P)-mediated endothelial cell (EC) barrier enhancement. Binds to NAA10/ARD1 and PTK2B/PYK2.

Similarity:

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family.

Research Fields

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 > Calcium signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Apelin signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.(View pathway)
· Organismal Systems > Immune system > Platelet activation.(View pathway)
· Organismal Systems > Digestive system > Gastric acid secretion.
· Organismal Systems > Circulatory system > Vascular smooth muscle contraction.(View pathway)
· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

WB Images(1)


ELISA analysis of AF8387 showing specificity to Phospho-MLCK (Tyr464) Blocking Peptide peptide. Peptides concentration: 1ug/ml.
P-peptide: phospho-peptide; N-peptide: non-phospho-peptide.
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Catalog Number :

AF8387-BP
(Blocking peptide available as AF8387-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Dog
67%
Zebrafish
0%
Chicken
0%
Rabbit
0%
Xenopus
0%
Pig
0%
Bovine
0%
Sheep
0%
Horse
0%
High similarity Medium similarity Low similarity No similarity
Q15746 as Substrate
Site PTM Type Enzyme
S13 Phosphorylation
S16 Phosphorylation
T88 Phosphorylation
Y104 Phosphorylation
T109 Phosphorylation
T118 Phosphorylation
S145 Phosphorylation
Y231 Phosphorylation P00519 (ABL1)
S287 Phosphorylation
S305 Phosphorylation
S324 Phosphorylation
S328 Phosphorylation
T331 Phosphorylation
T335 Phosphorylation
S343 Phosphorylation
S365 Phosphorylation
S367 Phosphorylation
S422 Phosphorylation
S460 Phosphorylation
Y464 Phosphorylation P00519 (ABL1) , P12931 (SRC)
S469 Phosphorylation
Y471 Phosphorylation P12931 (SRC)
T488 Phosphorylation
S515 Phosphorylation
Y556 Phosphorylation P00519 (ABL1)
K603 Methylation
K608 Acetylation
Y611 Phosphorylation P00519 (ABL1)
Y792 Phosphorylation P00519 (ABL1)
S827 Phosphorylation
Y846 Phosphorylation P00519 (ABL1)
S848 Phosphorylation
T883 Phosphorylation
S911 Phosphorylation
S947 Phosphorylation
T978 Phosphorylation
K1042 Acetylation
S1093 Phosphorylation
K1101 Methylation
K1103 Methylation
K1113 Acetylation
S1151 Phosphorylation
S1208 Phosphorylation Q13177 (PAK2)
S1209 Phosphorylation
T1230 Phosphorylation
Y1410 Phosphorylation
S1438 Phosphorylation
Y1449 Phosphorylation P00519 (ABL1)
K1496 Acetylation
Y1575 Phosphorylation P00519 (ABL1)
S1617 Phosphorylation
Y1635 Phosphorylation P00519 (ABL1)
K1744 Acetylation
K1747 Acetylation
T1748 Phosphorylation Q15746 (MYLK)
S1759 Phosphorylation Q13177 (PAK2)
S1760 Phosphorylation Q15746 (MYLK)
S1768 Phosphorylation Q15746 (MYLK)
S1772 Phosphorylation Q13153 (PAK1)
S1773 Phosphorylation P17612 (PRKACA)
T1774 Phosphorylation
S1776 Phosphorylation P49841 (GSK3B)
T1778 Phosphorylation
S1779 Phosphorylation P27361 (MAPK3) , P28482 (MAPK1)
S1788 Phosphorylation
S1793 Phosphorylation
S1824 Phosphorylation
Y1835 Phosphorylation
S1848 Phosphorylation
S1852 Phosphorylation
Q15746 as PTM Enzyme
Substrate Site Source
O14950 (MYL12B) T19 Uniprot
O14950 (MYL12B) S20 Uniprot
P10916 (MYL2) S15 Uniprot
P24844 (MYL9) T19 Uniprot
P24844 (MYL9) S20 Uniprot
Q05682 (CALD1) S73 Uniprot
Q05682 (CALD1) T666 Uniprot
Q05682 (CALD1) S700 Uniprot
Q05682 (CALD1) T735 Uniprot
Q05682 (CALD1) S744 Uniprot
Q15746 (MYLK) T1748 Uniprot
Q15746 (MYLK) S1760 Uniprot
Q15746 (MYLK) S1768 Uniprot
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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