Overview

  • Product name

    Anti-non-muscle Myosin IIA antibody [2B3]
    See all non-muscle Myosin IIA primary antibodies
  • Description

    Mouse monoclonal [2B3] to non-muscle Myosin IIA
  • Host species

    Mouse
  • Tested applications

    Suitable for: IP, WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment (GST-tag) corresponding to Human non-muscle Myosin IIA aa 1871-1960. MW of the GST tag alone is 26 KDa.
    Sequence:

    RLKQLKRQLEEAEEEAQRANASRRKLQRELEDATETADAMNREVSSLKNK LRRGDLPFVVPRRMARKGAGDGSDEEVDGKADGAEAKPAE


    Database link: P35579

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.4
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    2B3
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab55456 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
IP Use at an assay dependent concentration. PubMed: 22136066
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 215,250 kDa.
IHC-P Use a concentration of 0.5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function

    Cellular myosin that appears to play a role in cytokinesis, cell shape, and specialized functions such as secretion and capping.
  • Tissue specificity

    In the kidney, expressed in the glomeruli. Also expressed in leukocytes.
  • Involvement in disease

    Defects in MYH9 are the cause of May-Hegglin anomaly (MHA) [MIM:155100]. MHA is an autosomal dominant macrothrombocytopenia characterized by thrombocytopenia, giant platelets and leukokyte inclusions appearing as highly parallel paracrystalline bodies.
    Defects in MYH9 are the cause of Sebastian syndrome (SBS) [MIM:605249]. SBS is an autosomal dominant macrothrombocytopenia characterized by thrombocytopenia, giant platelets and leukocyte inclusions that are smaller and less organized than in May-Hegglin anomaly.
    Defects in MYH9 are the cause of Fechtner syndrome (FTNS) [MIM:153640]. FTNS is an autosomal dominant macrothrombocytopenia characterized by thrombocytopenia, giant platelets and leukocyte inclusions that are small and poorly organized. Additionally, FTNS is distinguished by Alport-like clinical features of sensorineural deafness, cataracts and nephritis.
    Defects in MYH9 are the cause of Alport syndrome with macrothrombocytopenia (APSM) [MIM:153650]. APSM is an autosomal dominant disorder characterized by the association of ocular lesions, sensorineural hearing loss and nephritis (Alport syndrome) with platelet defects.
    Defects in MYH9 are the cause of Epstein syndrome (EPS) [MIM:153650]. EPS is an autosomal dominant disorder characterized by the association of macrothrombocytopathy, sensorineural hearing loss and nephritis.
    Defects in MYH9 are the cause of deafness autosomal dominant type 17 (DFNA17) [MIM:603622]. DFNA17 is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information. DFNA17 is characterized by progressive hearing impairment and cochleosaccular degeneration.
    Defects in MYH9 are the cause of macrothrombocytopenia with progressive sensorineural deafness (MPSD) [MIM:600208]. MPSD is an autosomal dominant disorder characterized by the association of macrothrombocytopathy and progressive sensorineural hearing loss without renal dysfunction.
    Note=Subjects with mutations in the motor domain of MYH9 present with severe thrombocytopenia and develop nephritis and deafness before the age of 40 years, while those with mutations in the tail domain have a much lower risk of noncongenital complications and significantly higher platelet counts. The clinical course of patients with mutations in the four most frequently affected residues of MYH9 (responsible for 70% of MYH9-related cases) were evaluated. Mutations at residue 1933 do not induce kidney damage or cataracts and cause deafness only in the elderly, those in position 702 result in severe thrombocytopenia and produce nephritis and deafness at a juvenile age, while alterations at residue 1424 or 1841 result in intermediate clinical pictures.
    Note=Genetic variations in MYH9 are associated with non-diabetic end stage renal disease (ESRD).
  • Sequence similarities

    Contains 1 IQ domain.
    Contains 1 myosin head-like domain.
  • Domain

    The rodlike tail sequence is highly repetitive, showing cycles of a 28-residue repeat pattern composed of 4 heptapeptides, characteristic for alpha-helical coiled coils.
  • Post-translational
    modifications

    ISGylated.
  • Information by UniProt
  • Database links

  • Alternative names

    • BDPLT 6 antibody
    • BDPLT6 antibody
    • Cellular myosin heavy chain antibody
    • Cellular myosin heavy chain type A antibody
    • DFNA 17 antibody
    • DFNA17 antibody
    • EPSTS antibody
    • FTNS antibody
    • MGC104539 antibody
    • MHA antibody
    • MYH 2A antibody
    • MYH 9 antibody
    • MYH2A antibody
    • MYH9 antibody
    • MYH9_HUMAN antibody
    • MYHas8 antibody
    • MyHC 2A antibody
    • MyHC IIa antibody
    • MyHC2A antibody
    • MyHCIIa antibody
    • MYHSA 2 antibody
    • MYHSA2 antibody
    • Myosin 9 antibody
    • Myosin heavy chain 9 antibody
    • Myosin heavy chain 9 non muscle antibody
    • Myosin heavy chain antibody
    • Myosin heavy chain non muscle IIa antibody
    • Myosin heavy chain nonmuscle IIa antibody
    • Myosin heavy polypeptide 2 antibody
    • Myosin heavy polypeptide 9 non muscle antibody
    • Myosin-9 antibody
    • Myosin9 antibody
    • NMHC II A antibody
    • NMMHC A antibody
    • NMMHC II a antibody
    • NMMHC II-a antibody
    • NMMHC IIA antibody
    • NMMHC-A antibody
    • NMMHC-IIA antibody
    • NMMHCA antibody
    • Non muscle myosin heavy chain A antibody
    • Non muscle myosin heavy chain antibody
    • Non muscle myosin heavy chain II A antibody
    • Non muscle myosin heavy polypeptide 9 antibody
    • non-muscle IIa antibody
    • Non-muscle myosin heavy chain A antibody
    • Non-muscle myosin heavy chain IIa antibody
    • Nonmuscle myosin heavy chain A antibody
    • Nonmuscle myosin heavy chain II A antibody
    • type A antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: non-muscle Myosin IIA knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: HEK293 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab55456 observed at 230 kDa. Red - loading control, ab18251, observed at 52 kDa.

    ab55456 was shown to specifically react with non-muscle Myosin IIA when non-muscle Myosin IIA knockout samples were used. Wild-type and non-muscle Myosin IIA knockout samples were subjected to SDS-PAGE. ab55456 at a concentration of 1 µg/ml and ab18251 (loading control to alpha Tubulin) at a dilution of 1/1000 were incubated overnight at 4°C. Blots were developed withGoat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorecence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling non-muscle Myosin IIA (Green) using ab55456 at 10 µg/ml in ICC/IF analysis.

  • Immunofluorescence analysis of plakoglobin-knocked down MCF7 cells, staining non-muscle Myosin IIA with ab55456.

    Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked in 5% goat serum. Samples were incubated with primary antibody overnight at 4°C. An AlexaFluor®-conjugated anti-mouse IgG was used as the secondary antibody.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling non-muscle Myosin llA with ab55456 at 0.5 µg/ml.

  • Overlay histogram showing HEK293 cells stained with ab55456 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab55456, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • ICC/IF image of ab55456 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab55456, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Western blot against tagged recombinant protein immunogen using ab55456 non-muscle Myosin IIA antibody at 1ug/ml. Predicted band size of immunogen is 33 kDa
  • Anti-non-muscle Myosin IIA antibody [2B3] (ab55456) + HeLa
  • Anti-non-muscle Myosin IIA antibody [2B3] (ab55456) at 5 µg/ml + Human kidney tissue lysate

References

This product has been referenced in:

  • Zhang W  et al. Rho kinase collaborates with p21-activated kinase to regulate actin polymerization and contraction in airway smooth muscle. J Physiol 596:3617-3635 (2018). Read more (PubMed: 29746010) »
  • Dash B  et al. Multiple myosin motors interact with sodium/potassium-ATPase alpha 1 subunits. Mol Brain 11:45 (2018). Read more (PubMed: 30086768) »
See all 26 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Zebrafish Tissue lysate - whole (24 somite stage wild type whole embryos (deyolked))
Gel Running Conditions
Reduced Denaturing (7.5% SDS-PAGE 100 V)
Loading amount
25 µg
Specification
24 somite stage wild type whole embryos (deyolked)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 11 2016

Application
Immunoprecipitation
Sample
Zebrafish Tissue lysate - whole (24 somite stage wild type whole embryos (deyolked))
Total protein in input
40 µg
Immuno-precipitation step
Protein A/G
Specification
24 somite stage wild type whole embryos (deyolked)

Abcam user community

Verified customer

Submitted Nov 09 2016

Answer


Below is a link to our indirect flow cytometry protocol:
https://www.abcam.com/index.html?pageconfig=resource&rid=11381

If possible, I recommend not fixing your samples and analyzing them the same day as staining, as soon as possible after adding the secondary antibody.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Breast)
Specification
Breast
Fixative
Formaldehyde
Permeabilization
Yes - 0.5% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 15 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (YTS NK cell line)
Loading amount
200000 cells
Specification
YTS NK cell line
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris Gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 24°C

Ms. Keri Sanborn

Verified customer

Submitted Jul 25 2008

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