Overview

  • Product nameAnti-SM22 alpha antibody
    See all SM22 alpha primary antibodies
  • Description
    Goat polyclonal to SM22 alpha
  • Tested applicationsSuitable for: WB, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
    Predicted to work with: Cow, Dog
  • Immunogen

    Synthetic peptide: MTGYGRPRQIIS, corresponding to C terminal amino acids 189-200 of Human SM22 alpha.

  • Positive control
    • Hela cell, human muscle, heart, and placenta lysates.
  • General notes


    In contrast to fast and slow skeletal muscle cells that fuse and terminally differentiate, smooth muscle cells are able to simultaneously proliferate and express lineage-restricted proteins. One of these proteins, expressed exclusively in smooth muscles, has been referred to as SM22-alpha, a 22-kD protein with structural similarity to the vertebrate thin filament myofibrillar regulatory protein calponin and the Drosophila muscle protein mp20, neither of which play a direct role in the contractile apparatus. The protein is a transformation and shape-change sensitive actin cross-linking/gelling protein found in fibroblasts and smooth muscle. Its expression is down-regulated in many cell lines, and this down-regulation may be an early and sensitive marker for the onset of transformation. A functional role of this protein is unclear.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: 0.5% BSA, Tris-saline. pH 7.3
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Purification notesPurified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Primary antibody notesIn contrast to fast and slow skeletal muscle cells that fuse and terminally differentiate, smooth muscle cells are able to simultaneously proliferate and express lineage-restricted proteins. One of these proteins, expressed exclusively in smooth muscles, has been referred to as SM22-alpha, a 22-kD protein with structural similarity to the vertebrate thin filament myofibrillar regulatory protein calponin and the Drosophila muscle protein mp20, neither of which play a direct role in the contractile apparatus. The protein is a transformation and shape-change sensitive actin cross-linking/gelling protein found in fibroblasts and smooth muscle. Its expression is down-regulated in many cell lines, and this down-regulation may be an early and sensitive marker for the onset of transformation. A functional role of this protein is unclear.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab10135 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
WB Use a concentration of 0.01 - 0.03 µg/ml. Detects a band of approximately 22-23 kDa (predicted molecular weight: 23 kDa).Can be blocked with Human SM22 alpha peptide (ab23184).
Flow Cyt Use at an assay dependent concentration. PubMed: 17099777

ab37373 - Goat polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/50. PubMed: 16080196
ICC/IF Use a concentration of 5 µg/ml.

Target

Anti-SM22 alpha antibody images



  • Predicted band size : 23 kDa

    ab10135 staining (0.2µg/ml) of Human Muscle lysate (RIPA buffer, 35µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence.

    ab10135 staining (0.2µg/ml) of Human Muscle lysate (RIPA buffer, 35µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.
  • Immunofluorescence analysis of REF52 (rat embryo fibroblast) cells, staining SM22 alpha (green) with ab10135.

    Cells were fixed with paraformaldehyde for 10 minutes and permeabilized with 0.05% Triton X-100, before incubating with primary antibody. A fluorescent-conjugated anti-goat IgG was used as the secondary antibody.
  • Anti-SM22 alpha antibody (ab10135) at 0.5 µg/ml + Human muscle at 20 µg

    Secondary
    Donkey Anti-Goat IgG H&L (HRP) (ab6885) at 1/5000 dilution

    Predicted band size : 23 kDa
    Observed band size : 24 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa,37 kDa,42 kDa,50 kDa,75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 5 minutes
    The band at 50kDa is recognised as contamination.
  • ICC/IF image of ab10135 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal donkey 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 (ab10135, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat 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.

  • Anti-SM22 alpha antibody (ab10135) at 0.02 µg/ml + Human Placenta lysate at 35 µg

    Predicted band size : 23 kDa
    Observed band size : 23 kDa
    Primary incubation was 1 hour. Detected by chemiluminescence.
  • Immunohistochemical analysis of PFA-fixed, paraffin embedded murine kidney tissue, staining SM22 alpha with ab10135 at 1/100 dilution.

References for Anti-SM22 alpha antibody (ab10135)

This product has been referenced in:
  • Ahnstedt H  et al. U0126 attenuates cerebral vasoconstriction and improves long-term neurologic outcome after stroke in female rats. J Cereb Blood Flow Metab 35:454-60 (2015). Read more (PubMed: 25492115) »
  • Manderfield LJ  et al. Hippo signaling is required for Notch-dependent smooth muscle differentiation of neural crest. Development 142:2962-71 (2015). IHC-P ; Mouse . Read more (PubMed: 26253400) »

See all 29 Publications for this product

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Because antibodies are immunoglobins you would have to find concentration by measuring those proteins. However, because ab28811 is a tissue culture supernate it would neccessarily contain other proteins that th...

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Thank you for your email and I'm sorry to hear that you are experiencing some difficulty with these antibodies. All 3 of these antibodies have been successfully tested for application in Western blotting. However, these antibodies have not yet been...

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Thank you for your email. Regarding ab14106 and ab10135 (the SM22 alpha antibodies), both are guaranteed to work in Western blotting with human samples. I can't say which one works better, and suggest that you take a closer look at the online datasheet...

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Thank you for your enquiry. We have the following SM22 alpha antibodies, ab14106 and ab10135. Yes, both work in Western blotting and although we don't have any information regarding the antibody detection limits, there are Western blotting images av...

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Application Western blot
Sample Dog Tissue lysate - whole (heart valve)
Specification heart valve
Blocking step Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: n/a
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Verified customer

Submitted Aug 15 2005

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (heart)
Specification heart
Blocking step Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: N/A
Username

Abcam user community

Verified customer

Submitted Aug 12 2005

Thank you for your enquiry regarding ab10135. To our knowledge, this antibody has yet to be tested in IHC. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how yo...

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Thank you for your enquiry. If we understand this query, you are asking whether the antibody binds an epitope on the cell surface which will allow them to sort cells using magnetic beads. According to Swiss-Prot - SM22 alpha looks like a cytoplasmic...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"