Overview

  • Product name
  • Description
    Goat polyclonal to SM22 alpha
  • Host species
    Goat
  • Tested applications
    Suitable for: WB, Flow Cyt, IHC-Fr, IHC-P, IHC-FoFr, 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
    • WB: Human muscle, heart, and placenta lysates. ICC/IF: REF52 and HeLa cells IHC-P: Human Uterus tissue, murine kidney tissue
  • 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

  • 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
    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Primary antibody 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.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

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-Fr Use at an assay dependent concentration. PubMed: 28955046
IHC-P 1/50. PubMed: 16080196
IHC-FoFr Use at an assay dependent concentration. PubMed: 26183159
ICC/IF Use a concentration of 5 µg/ml. PubMed: 22257561

Target

Images

  • ab10135 (5µg/ml) staining of paraffin embedded Human Uterus. Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
  • Anti-SM22 alpha antibody (ab10135) at 0.03 µg + Human Heart lysate at 1/35 dilution

    Predicted band size: 23 kDa

  • 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.
  • Immunohistochemical analysis of PFA-fixed, paraffin embedded murine kidney tissue, staining SM22 alpha with ab10135 at 1/100 dilution.
  • 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.

  • 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.
  • 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.

References

This product has been referenced in:
  • Neumann P  et al. The lncRNA GATA6-AS epigenetically regulates endothelial gene expression via interaction with LOXL2. Nat Commun 9:237 (2018). Read more (PubMed: 29339785) »
  • Wang J  et al. Tfap2b mutation in mice results in patent ductus arteriosus and renal malformation. J Surg Res 227:178-185 (2018). Read more (PubMed: 29804851) »
See all 67 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer

Thank you for contacting us.

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 the cell would produce or might be in the media. Therefore any protein measurements that you could make would not be a particularly accurate measure of the antibody present. I am sorry that I could not provide a better solution for you.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

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 tested for application in IHC to our knowledge. They very well may not be suitable for this particular application. I do suggest that you submit Abreviews regarding your experiences with these antibodies in IHC as your feedback is extremely useful for us as well as other customers. To submit an Abreview, just click on the Abreviews tab located on the online product datasheet and then click on the submit an Abreview link. In exchange for an Abreview we will award 50 Abpoints (an additional 100 Abpoints if an image is submitted) which can be exchanged for a variety of awards. In order to check to see if your secondary antibody is causing the background that you are seeing, you can test this by omitting primary antibody in your protocol and testing with secondary antibody only. If background appears, change either the secondary antibody or the blocking agent. At the bottom of the online datasheets for these antibodies are a list of compatible secondaries (such as ab6741) which are suitable for use with these primary antibodies. For ab1301 (Goat polyclonal to LASP1), an approximate 30 kDa band was seen in Mouse Brain extracts. I would therefore suggest using this as a positive control. I hope this helps. Please don't hesitate to contact us again if you need additional assistance with these antibodies. Happy New Year!

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Answer

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 datasheets before choosing the one that best suits your needs. We have one antibody to smooth muscle actin (ab21027) and this antibody is also guaranteed to work in Western blotting with human samples. If you need an alpha smooth muscle actin antibody instead, please take a look at products ab15734, ab5694 and ab7817, for example. I hope this helps. Please contact us again if you have any additional questions.

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Answer

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 available on the online datasheets. Also, what species are you working on? This will help you to narrow your choice as well. For smooth muscle actin, we have ab21027 - Goat polyclonal to smooth muscle Actin - and this antibody works in Western blotting with human samples. There is a Western blot image available on the online datasheet. We also have some alpha smooth muscle actin antibodies that work in Western blotting: ab15734, ab5694 and ab7817 for example. If you have any additional questions, please contact us again.

Read More
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

Abcam user community

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

Abcam user community

Verified customer

Submitted Aug 12 2005

Answer

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 you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher. Please contact us again if you need further assistance.

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Answer

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 protein. Therefore we would suggestion trying to get the antibody working in flow cytometry (we haven't tried this) and then sort them in a FACSorter.

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