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
  • Host species
    Mouse
  • 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.

  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. 

Properties

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

    All lanes : Human vascular smooth muscle whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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


    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:
  • Fredriksson-Lidman K  et al. Sorbin and SH3 domain-containing protein 2 (SORBS2) is a component of the acto-myosin ring at the apical junctional complex in epithelial cells. PLoS One 12:e0185448 (2017). Read more (PubMed: 28961272) »
  • Ferrán B  et al. The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation. Sci Rep 6:25944 (2016). WB . Read more (PubMed: 27181368) »
See all 27 Publications for this product

Customer reviews and Q&As

1-10 of 26 Abreviews or Q&A

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

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

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

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

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

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

Miss. Cristina Rodriguez

Verified customer

Submitted Apr 12 2013

Answer

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 diesem Produkt enthalten sind, nicht nur den Antikörper.

2. Ascites hat eine recht geringe, und schwankende, Antikörperkonzentration: Wenn dieser Antikörper aufgereinigt wird, bleibt am Ende recht wenig übrig, was konjugiert werden kann.

3. Wenn Sie einen aufgereinigten Antikörper konjugieren wollen, brauchen Sie normalerweise eine Minimum- Konzentration, dh, ein aufgereinigter Ascites Antikörper müsste daher auch noch aufkonzentriert werden.

Zwar bieten wir für alle diese Schritte ein entsprechendes Kit an, die Kosten würden allerdings dementsprechend steigen. Noch teurer würde es werden, falls wir dies in unserem Labor durchführen.

Muss es den unbedingt dieser Antikörper sein, oder würde auch ein aufgereinigter polyklonaler Kaninchen Antikörper für Sie in frage kommen?

Read More

Answer

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 in Mouse tissues. However, in order to avoid cross reactivity and improve the signal intensity we recommend using the Mouse on Mouse Polymer IHC Kit: https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html or if available, using serum from the secondary species to block unspecific signals.

We recommend using a secondary raised in a species such as goat or donkey. The https://www.abcam.com/index.html?pageconfig=resource&rid=11465 are also helpful in order to perform a Mouse on Mouse staining procedure.

I hope this information was helpful. Please do not hesitate to contact us for further assistance.

Have a great day.

Read More

Question
Answer

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 only just received a reply.

I am sorry to confirm that the concentration has indeed not been detemined. As discussed on the telephone, I can confirm that ab684 is sold as ascites fluid. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not normally have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for ascites, concentration of antibody is estimated to vary between 5 - 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you.

Regarding concentraiton to use in IHC-P, I am unfortunately not sure how the concentration to use was determined at 4 ug/ml. I can suggest that the dilution used will require some individual optimization and can recommend a titration experiment using several dilutions to determine which is best.

If you require anything further, please do not hesitate to let me know.

Read More

Answer

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.

Concerning our testing discount, you are not obligated to test ab684 in IP if you decide not to over the course of your research. However, please remember that if you do wish to test this antibody in IP you must submit your results to us before the expiration date of the code which is valid for 4 months after the date at which it is issued. The details for this testing discount are below:

DISCOUNT CODE:xxxxxxxxxx
Expiration date: xxxxxxxxxxxx
Value: €xxxxxxxx

This code will give you €xxxxxxx off your next order before the expiration date. To redeem this offer, please submit an Abreview for IP and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be used to purchase any other Abcam product. The code can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/abtrial.

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1-10 of 26 Abreviews or Q&A

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