Recombinant
RabMAb

Recombinant Anti-Myosin light chain kinase/MLCK antibody [EP1458Y] (ab76092)

Overview

  • Product name

    Anti-Myosin light chain kinase/MLCK antibody [EP1458Y]
    See all Myosin light chain kinase/MLCK primary antibodies
  • Description

    Rabbit monoclonal [EP1458Y] to Myosin light chain kinase/MLCK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Myosin light chain kinase/MLCK aa 1850-1950 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Human brain lysate IHC-P: Human smooth muscle tissue IF: SH-SY5Y cells
  • General notes

    Previously labeled as Myosin light chain kinase.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP1458Y
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab76092 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/5000 - 1/10000. Predicted molecular weight: 211 kDa.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Use of HRP-conjugated or polymerized HRP secondary antibodies, stronger signals have been found using the polymerized HRP secondary.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Calcium/calmodulin-dependent enzyme implicated in smooth muscle contraction via phosphorylation of myosin light chains (MLC). Also regulates actin-myosin interaction through a non-kinase activty (By similarity). Implicated in the regulation of endothelial as well as vascular permeability. 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.
    • 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.
    • Sequence similarities

      Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family.
      Contains 1 fibronectin type-III domain.
      Contains 9 Ig-like C2-type (immunoglobulin-like) domains.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications

      MLCK is probably down-regulated by phosphorylation.
      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.
    • Information by UniProt
    • Database links

    • Alternative names

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

    Images

    • ab76092 staining Myosin light chain kinase/MLCK in mouse oocyte cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 overnight at 4°C and blocked with 3% BSA for 12 hours at 4°C. Samples were incubated with primary antibody (1/50) for 12 hours at 4°C. A Cy5®-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

      See Abreview

    • Immunohistochemical analysis of paraffin embedded human smooth muscle, using ab76092, at 1/100 dilution.

      Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Turner SR  et al. Smoothelin-like 1 deletion enhances myogenic reactivity of mesenteric arteries with alterations in PKC and myosin phosphatase signaling. Sci Rep 9:481 (2019). Read more (PubMed: 30679490) »
    • Wang J  et al. MicroRNA-92a promotes vascular smooth muscle cell proliferation and migration through the ROCK/MLCK signalling pathway. J Cell Mol Med 23:3696-3710 (2019). Read more (PubMed: 30907506) »
    See all 28 Publications for this product

    Customer reviews and Q&As

    1-10 of 19 Abreviews or Q&A

    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (MDCK)
    Gel Running Conditions
    Non-reduced Denaturing
    Loading amount
    20 µg
    Treatment
    siMLCK
    Specification
    MDCK
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Jan 03 2019

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Prostate)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate, pH 6
    Permeabilization
    No
    Specification
    Prostate
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Nov 28 2017

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Embryonic Heart)
    Permeabilization
    Yes - Tween-20
    Specification
    Embryonic Heart
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 31 2016

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Heart)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    30 µg
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 20 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (colon from Nonalcoholic Steatohepatitis mouse)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 0.5% typsin
    Specification
    colon from Nonalcoholic Steatohepatitis mouse
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 22 2015

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Intestinal tissue)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis Tris MOPS)
    Loading amount
    28 µg
    Specification
    Intestinal tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 16 2015

    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Intestinal tissue)
    Gel Running Conditions
    Reduced Denaturing (4-12% BIS TRIS)
    Loading amount
    20 µg
    Specification
    Intestinal tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 16 2015

    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Intestinal tissue)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis Tris MOPS)
    Loading amount
    28 µg
    Specification
    Intestinal tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Sep 21 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Oocyte)
    Permeabilization
    Yes - 0.1%triton 4? overnight
    Specification
    Oocyte
    Blocking step
    BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 12 2015

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human arterial myocytes)
    Permeabilization
    Yes - 0.3% Triton X-100
    Specification
    Human arterial myocytes
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Apr 08 2015

    1-10 of 19 Abreviews or Q&A

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