Anti-non-muscle Myosin IIB antibody [3H2] (ab684)

Overview

  • Product name
    Anti-non-muscle Myosin IIB antibody [3H2]
    See all non-muscle Myosin IIB primary antibodies
  • Description
    Mouse monoclonal [3H2] to non-muscle Myosin IIB
  • Specificity
    This antibody reacts with non-muscle myosin heavy chain (SMemb).
  • Tested applications
    Suitable for: IHC-P, ICC/IF, WB, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Hamster, Human
  • Immunogen

    Synthetic peptide:

    TSDVNETQPPQSE

    , corresponding to C (beta) terminal amino acids 1964-1976 of Human non-muscle Myosin IIB.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Ascitic fluid
  • Concentration information loading...
  • Purity
    Ascites
  • Clonality
    Monoclonal
  • Clone number
    3H2
  • Isotype
    IgG2b
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab684 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
IHC-P 1/100.
ICC/IF Use at an assay dependent concentration. PubMed: 17478566Use at an assay dependent dilution.
WB 1/2000.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

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

Target

  • Function
    Cellular myosin that appears to play a role in cytokinesis, cell shape, and specialized functions such as secretion and capping. Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.
  • Tissue specificity
    Isoform 1 is expressed in cerebellum and spinal chord. Isoform 2 is expressed in cerebrum and retina. Isoform 3 is expressed in the cerebrum and to a much lower extent in cerebellum.
  • 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
    Phosphorylated by ABL2.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cellular myosin heavy chain antibody
    • Cellular myosin heavy chain type B antibody
    • Cellular myosin heavy chain type B type B antibody
    • MGC134913 antibody
    • MGC134914 antibody
    • MYH 10 antibody
    • Myh10 antibody
    • MYH10_HUMAN antibody
    • Myosin 10 antibody
    • Myosin heavy chain 10 antibody
    • Myosin heavy chain 10 non muscle antibody
    • Myosin heavy chain antibody
    • Myosin heavy chain non muscle 11b antibody
    • Myosin heavy chain nonmuscle IIb antibody
    • Myosin heavy chain nonmuscle type B antibody
    • Myosin heavy polypeptide 10 non muscle antibody
    • Myosin-10 antibody
    • Myosin10 antibody
    • NMMHC B antibody
    • NMMHC II b antibody
    • NMMHC II-b antibody
    • NMMHC IIB antibody
    • NMMHC-B antibody
    • NMMHC-IIB antibody
    • NMMHCB antibody
    • Non muscle myosin heavy chain B antibody
    • Non muscle myosin heavy chain IIB antibody
    • Non muscle myosin II heavy chain B antibody
    • non-muscle IIb antibody
    • Non-muscle myosin heavy chain B antibody
    • Non-muscle myosin heavy chain IIb antibody
    • Nonmuscle myosin heavy chain B antibody
    • Nonmuscle myosin heavy chain IIB antibody
    • Nonmuscle myosin II heavy chain B antibody
    • Nonmuscle myosin IIB antibody
    • type B antibody
    see all

Images

  • All lanes : Anti-non-muscle Myosin IIB antibody [3H2] (ab684) at 1/2000 dilution

    Lane 1 : Human vascular smooth muscle whole cell lysate
    Lane 2 : Human vascular smooth muscle whole cell lysate
    Lane 3 : Human vascular smooth muscle whole cell lysate
    Lane 4 : Human vascular smooth muscle whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated sheep anti-mouse IgG at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size : 200 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 1 hour at 20°C.

    Incubated with the primary antibody in 5% milk for 16 hours at 4°C.

    The 4 lanes were each loaded with 10ug of protein based upon a BCA assay for protein quantification. However, looking at a subsequent loading control blot (vinvulin) lanes 1 and 4 are under loaded versus lanes 2 and 3.

    See Abreview

  • ab684 staining non-muscle Myosin IIB in WIF-B cells (hepatoma-derived hybrid cell line) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1% donkey serum in 1% PBST for 1 hour at 22°C. Samples were incubated with primary antibody (1/50 in blocking buffer) for 3 hours at 22°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Ab684 staining nnon-muscle Myosin IIB in Human colon.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • Overlay histogram showing HeLa cells stained with ab684 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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. The cells were then incubated with the antibody (ab684, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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 HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

References

This product has been referenced in:
  • Feng J  et al. Depletion of kinesin-12, a myosin-IIB-interacting protein, promotes migration of cortical astrocytes. J Cell Sci 129:2438-47 (2016). Read more (PubMed: 27170353) »
  • Sur S  et al. Increased Expression of Phosphorylated Polo-Like Kinase 1 and Histone in Bypass Vein Graft and Coronary Arteries following Angioplasty. PLoS One 11:e0147937 (2016). IF . Read more (PubMed: 26820885) »

See all 26 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (HepG2 Cells)
Gel Running Conditions
Reduced Denaturing (8% SDS-PAGE)
Loading amount
20 µg
Specification
HepG2 Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Armen Petrosyan

Verified customer

Submitted Dec 21 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Dec 21 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (WIF-B cells, the hepatoma-derived hybrid cell line)
Permeabilization
Yes - 0.2% Triton X-100
Specification
WIF-B cells, the hepatoma-derived hybrid cell line
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

Submitted Dec 08 2016

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (4-12% SDS-PAGE)
Sample
Human Cell lysate - other (293T)
Specification
293T
Treatment
siRNA transfected for 2 days
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 15 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Aorta)
Specification
Aorta
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate pH6
Permeabilization
No
Blocking step
Blocking reagent from ab127055 kit as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Username

Miss. Cristina Rodriguez

Verified customer

Submitted Apr 19 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (VSMC)
Loading amount
25 µg
Specification
VSMC
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Miss. Cristina Rodriguez

Verified customer

Submitted Apr 12 2013

Leider bieten wir diesen Antikörper nicht in der konjugierten From an. Dies hat mehrere Gründe:

1. Es handelt sich um ein Ascites Produkt: Wenn Sie dieses Produkt konjugieren, so wie es ist, konjugieren Sie ALLE Proteine, die in dies...

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Thank you for contacting us.

I am afraid the only alternative available against this target previously tested in Mouse tissues is ab684.

This antibody has successfully been tested (and is therefore guaranteed) to target this protein...

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Thank you for your recent telephone enquiry and for your patience.

I can confirm that this particular product is sourced externally. I have been in contact with them in order to obtain this information for you urgently, and regrettably have ...

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Thank you for contacting us.

My source of information for finding the isoform data for this protein was indeed uniprot. See: http://www.uniprot.org/uniprot/P35580

I can also confirm that this antibody is guarenteed to work in mouse....

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