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

概述

  • 产品名称Anti-Myosin light chain (phospho S20)抗体
    参阅全部 Myosin light chain 一抗
  • 描述
    兔多克隆抗体to Myosin light chain (phospho S20)
  • 特异性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.
  • 经测试应用适用于: ICC/IF, ELISA, WB, IHC-FoFr, IP, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Caenorhabditis elegans
    预测可用于: Bird, all Mammals不与反应: Xenopus laevis
  • 免疫原

    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).

  • 阳性对照
    • Cardiac myocytes.

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液Preservative: 0.01% Sodium Azide
    Constituents: 0.15M Sodium chloride, 0.02M Potassium phosphate, pH 7.2
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 纯化说明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.
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能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.
  • 组织特异性Smooth muscle tissues and in some, but not all, nonmuscle cells.
  • 序列相似性Contains 3 EF-hand domains.
  • 翻译后修饰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
  • 数据库链接
  • 别名
    • 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

Anti-Myosin light chain (phospho S20) antibody 图像



  • Predicted band size : 20 kDa

    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.

Anti-Myosin light chain (phospho S20) antibody (ab2480)参考文献

This product has been referenced in:
  • Wagstaff L  et al. Mechanical cell competition kills cells via induction of lethal p53 levels. Nat Commun 7:11373 (2016). ICC/IF . Read more (PubMed: 27109213) »
  • Ossipova O  et al. Vangl2 cooperates with Rab11 and Myosin V to regulate apical constriction during vertebrate gastrulation. Development 142:99-107 (2015). IHC-Fr ; Xenopus laevis . Read more (PubMed: 25480917) »

See all 23 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions Reduced Denaturing (4-12% BIS TRIS)
Loading amount 15 µg
Specification Intestinal tissue
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Sep 08 2015

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

Abcam user community

Verified customer

提交于 Apr 08 2015

Western blot

Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (15)
Sample Human Tissue lysate - whole (Heart)
Specification Heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 22 2013

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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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - other (Heart)
Loading amount 20 µg
Specification Heart
Gel Running Conditions Reduced Denaturing (15)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jun 07 2012

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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