Product nameAnti-non-muscle Myosin IIB antibody [3H2]
See all non-muscle Myosin IIB primary antibodies
DescriptionMouse monoclonal [3H2] to non-muscle Myosin IIB
SpecificityThis antibody reacts with non-muscle myosin heavy chain (SMemb).
Tested applicationsSuitable for: IHC-P, ICC/IF, WB, IHC-Fr, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Rabbit, Hamster, Human
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Concentration information loading...
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.
|ICC/IF||Use at an assay dependent concentration. PubMed: 17478566Use at an assay dependent dilution.|
|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.
FunctionCellular 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 specificityIsoform 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 similaritiesContains 1 IQ domain.
Contains 1 myosin head-like domain.
DomainThe rodlike tail sequence is highly repetitive, showing cycles of a 28-residue repeat pattern composed of 4 heptapeptides, characteristic for alpha-helical coiled coils.
modificationsPhosphorylated by ABL2.
- Information by UniProt
- Cellular myosin heavy chain antibody
- Cellular myosin heavy chain type B antibody
- Cellular myosin heavy chain type B type B antibody
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.
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.
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.
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.
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) »