Anti-Myosin light chain (phospho S20) antibody (ab2480)


  • Product name
    Anti-Myosin light chain (phospho S20) antibody
    See all Myosin light chain primary antibodies
  • Description
    Rabbit polyclonal to Myosin light chain (phospho S20)
  • Host species
  • Specificity
    This phospho specific polyclonal antibody is specific for phosphorylated pS20 of human myosin light chain. Reactivity with non-phosphorylated human myosin light chain is less than 1% by ELISA. The selected peptide sequence used to generate the polyclonal antibody is located near the amino terminal end of the polypeptide corresponding to the smooth/non-muscle form of myosin regulatory light chain found in cardiac myocytes in addition to smooth and non-muscle cells. This sequence differs from that of the sarcomeric/cardiac form of myosin regulatory light chain that has a different sequence around the phosphorylation site. BLAST search analysis was used to determine that the smooth and non-muscle forms of myosin regulatory light chain have identical sequences. Cross reactivity is expected.
  • Tested applications
    Suitable for: ICC/IF, ELISA, WB, IHC-FoFr, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Caenorhabditis elegans
    Predicted to work with: Bird, MammalsDoes not react with: Xenopus laevis
  • Immunogen

    Human Myosin Light Chain phospho peptide corresponding to a region of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH) - KKRPQRAT(pS)NVFAMFD (aa 12-27).

  • Positive control
    • Cardiac myocytes.


  • Form
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    This antibody was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide.
  • Clonality
  • Isotype
  • Research areas


Our Abpromise guarantee covers the use of ab2480 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
ICC/IF Use a concentration of 2.5 µg/ml.
ELISA 1/5000 - 1/20000.
WB 1/1000 - 1/5000. Predicted molecular weight: 20 kDa. We recommend using BSA for blocking when using phospho-specific antibodies.
IHC-FoFr Use at an assay dependent concentration. PubMed: 19279134

Trichloroacetic acid was used for tissue fixation.

IP 1/100.
IHC-P Use at an assay dependent concentration.


  • Function
    Myosin regulatory subunit that plays an important role in regulation of both smooth muscle and nonmuscle cell contractile activity via its phosphorylation. Implicated in cytokinesis, receptor capping, and cell locomotion.
  • Tissue specificity
    Smooth muscle tissues and in some, but not all, nonmuscle cells.
  • Sequence similarities
    Contains 3 EF-hand domains.
  • Post-translational
    Phosphorylation increases the actin-activated myosin ATPase activity and thereby regulates the contractile activity. It is required to generate the driving force in the migration of the cells but not necessary for localization of myosin-2 at the leading edge.
  • Information by UniProt
  • Database links
  • Alternative names
    • 20 kDa myosin light chain antibody
    • KMLC antibody
    • LC20 antibody
    • MLC-2C antibody
    • MLCK2 antibody
    • MRLC1 antibody
    • MYL9 antibody
    • MYL9_HUMAN antibody
    • Myosin regulatory light chain 2 antibody
    • Myosin regulatory light chain 2, smooth muscle isoform antibody
    • Myosin regulatory light chain 9 antibody
    • Myosin regulatory light chain MRLC1 antibody
    • Myosin regulatory light polypeptide 9 antibody
    • Myosin RLC antibody
    • Myosin, light chain 9, regulatory antibody
    • MYRL2 antibody
    • smooth muscle isoform antibody
    see all


  • Rabbit polyclonal to phospho Myosin Light Chain phospho Myosin Light Chain (Ser 20) (ab2480) used at a 1/5000 dilution to detect myosin light chain by Western blot. Either 13 ug (lanes 1-3) or 20 ug (lanes 4-7) of a mouse cardiac myocyte lysate was loaded on a 4-20% Criterion gel for SDS-PAGE.  Samples were either mock-treated or CLA treated:
    Lane 1 : untreated 45 min
    Lane 2 : CLA 50 nm 45 min
    Lane 3 : CLA 100 nm 45 min
    Lane 4 : A21 untreated 45 min
    Lane 5 : A22 A23187 5 min
    Lane 6 : A23 A23187 15 min
    Lane 7 : A24 A23187 60 min
    After washing, a 1/5,000 dilution of anti-rabbit HRP (ab7090) was used as secondary.

    Rabbit polyclonal to phospho Myosin Light Chain phospho Myosin Light Chain (Ser 20) (ab2480) used at a 1/5000 dilution to detect myosin light chain by Western blot. Either 13 ug (lanes 1-3) or 20 ug (lanes 4-7) of a mouse cardiac myocyte lysate was loaded on a 4-20

  • IHC-P of ab2480 at 2.5 µg/ml staining both vascular and myometrial smooth muscle cells of the uterus.  The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.

  • ab2480 staining Myosin light chain (phospho S20) in Xenopus laevis embryos by immunohistochemistry (PFA-perfusion fixed frozen tissue sections). The tissue sections were fixed in 2% TCA for 30 minutes and washed with PBST (PBS+0.3% TritonX 100) for 30 minutes. Samples were then blocked in 10% normal goat serum (NGS) for 1 hour at room temperature and incubated with the primary antibody. A Cy5 labelled Goat polyclonal to rabbit IgG was used as secondary.

    The arrows shows that depleting E-cadherin does not have any effect on the expression of apical pMLC.

  • Immunofluroescence analysis of Nicaragua cichlid (Hypsophrys nicaraguensis) keratocytes, staining Myosin light chain (phospho S20) (green) with ab2480.

    Cells were cultured on intermediate adhesion strength surfaces and extracted with 4% PEG and 1% Triton X-100 in cytoskeleton stabilizing buffer. Cells were blocked with PBS-BT for 5 minutes, and then incubated with primary antibody diluted in PBS-BT. Cells were then fixed with 4% formaldehyde in PBS for 10 minutes.


This product has been referenced in:
  • Cervero P  et al. Lymphocyte-specific protein 1 regulates mechanosensory oscillation of podosomes and actin isoform-based actomyosin symmetry breaking. Nat Commun 9:515 (2018). Read more (PubMed: 29410425) »
  • Shinoda T  et al. Elasticity-based boosting of neuroepithelial nucleokinesis via indirect energy transfer from mother to daughter. PLoS Biol 16:e2004426 (2018). Read more (PubMed: 29677184) »

See all 51 Publications for this product

Customer reviews and Q&As

Thank you for contacting us.

Are you looking for a specific light chain (i.e. 2 or 3)? We only have 2 other anti-myosin light chain antibodies as ab97891 and ab88726 but we don't have any WB images showing 2 bands for phopsho and unphosphory...

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You are correct that Abcam has not specifically tested cross reactivity of ab2480 with skeletal muscle isoform MLC. This cross reactivity is predicted based on similarity of the immunogen sequence to skeletal muscle MLC. Likewise, ab2480 has ...

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There are some notes regarding cross reactivity of this antibody under the Specificity section of the online datasheet. The notes state that cross reactivity of this antibody to smooth and non-muscle forms of myosin regulatory light chain should be det...

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Thank you for contacting Abcam.

The concentration of ab2480 is available on our public website and the concentration of the current lot that we have is 0.92mg/ml.

As to how many western blot or IHC tests that you would get out of vi...

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Hello and thank you for your patience in receiving this response. I was waiting to hear back from the laboratory which is based in the UK. IHC-P was tested, as the image indicates, and has now been added as a tested application to our datasheet. This w...

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Thank you for the information you provided about your experience with our myosin light chain pS19/20 antibody, ab2480. I have issued a free of charge replacement with the order number ***** for the delta catenin antibody, ab64239. To check the st...

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Thank you for the extra details. I look forward to hearing back from you about the results of your tests.

Thank you for sending the details of your protocol. As I mentioned in a previous e-mail, your protocol is similar to that of the one used for the green fluorescent image of Xenopus embryos, except the tissue was first fixed with PFA, I am not sure i...

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Thank you for contacting us about the problems you are having with these antibodies and IHC. I think the protocol you are using is similar to one that is shown on the datasheet for the phospho S20 antibody ab2480, under the green IF image of Xenopus em...

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Thank you for your enquiry. Regarding ab2480 (Myosin light chain (phospho S20) antibody), this antibody recognizes the phosphorylated serine in the immunogenic peptide "KKRPQRAT(pS)NVFAMFD". When I blasted this immunogenic peptide sequence in BLAST...

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