Overview

  • Product name
  • Description
    Rabbit polyclonal to SM22 alpha
  • Host species
    Rabbit
  • Specificity
    Recognises a band at 23kD that is specifically blocked by the immunising peptide.
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IHC-Fr, WB, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human, Pig
  • Immunogen

    Synthetic peptide within Mouse SM22 alpha aa 150 to the C-terminus conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab16067)

  • Positive control
    • WB: Human smooth muscle cell lysate. HeLa whole cell and nuclear extract. Human skeletal muscle tissue lysate. Human, mouse and rat colon tissue lysate. ICC/IF: HeLa cells.
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab14106 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
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC-P 1/200.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
ICC Use at an assay dependent concentration.

Target

Images

  • All lanes : Anti-SM22 alpha antibody (ab14106) at 1/1000 dilution

    All lanes : Lysates prepared from human primary smooth muscle cells

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated bovine polyclonal to rabbit IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa


    Exposure time: 2 minutes

    See Abreview

  • ab14106, at 1/100, staining SM22 alpha in chicken smooth muscle tissue sections by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections).

    Sections were fixed in methacarn (60% methanol, 30% chloroform, 10% acetic acid), prior to blocking in 2% serum for 5 minutes at 25°C and then incubated with ab14106, for 1 hour at 25°C. An Alexa-Fluor® 488 goat polyclonal to rabbit Ig, diluted 1/100, was used as the secondary antibody.

    See Abreview

  • ab14106 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed in 100% methanol for 5 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14106 at 1 µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature.

    DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1 hour at room temperature.

  • ab14106 staining SM22 alpha in mouse muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/1000.

    See Abreview

  • All lanes : Anti-SM22 alpha antibody (ab14106) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa nuclear extract
    Lane 3 : Human skeletal muscle tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Predicted band size: 23 kDa
    Observed band size: 23 kDa



    The band was completed abolished by peptide blocking with ab16067 (immunizing peptide) - not shown.

     


     

     

     

  • All lanes : Anti-SM22 alpha antibody (ab14106) at 1 µg/ml

    Lane 1 : Human colon tissue lysate - total protein (ab30051)
    Lane 2 : Mouse colon tissue lysate
    Lane 3 : Rat colon tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 23 kDa
    Additional bands at: 115 kDa, 20 kDa, 36 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab14106 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Anti-SM22 alpha antibody (ab14106) at 1 µg/ml + Recombinant Human SM22 alpha protein (ab101469) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa


    Exposure time: 30 seconds

References

This product has been referenced in:
See all 171 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-10 of 26 Abreviews

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse aortic smooth muscle cells)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
25 µg
Treatment
Cre or GFP adenovirus 6 hrs
Specification
Mouse aortic smooth muscle cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (vascular smooth muscle cells)
Permeabilization
No
Specification
vascular smooth muscle cells
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Carotid artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: sodium citrate solution
Permeabilization
Yes - 0.3% Triton X-100/PBS
Specification
Carotid artery
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
IHC - Wholemount
Sample
Mouse Tissue (artery)
Specification
artery

Abcam user community

Verified customer

Submitted Mar 01 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Adult Heart)
Permeabilization
Yes - 0.5% Tween-20 in block
Specification
Adult Heart
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 03 2017

Application
Western blot
Sample
Cow Tissue lysate - whole (Kidney)
Gel Running Conditions
Reduced Denaturing (14)
Loading amount
20 µg
Specification
Kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Sep 28 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
vascular smooth muscle cell
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted May 05 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human arterial myocytes)
Specification
Human arterial myocytes
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 08 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Cell (muscle cell)
Specification
muscle cell
Permeabilization
Yes - 0.1%Triton X-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
PBS, 1% BSA, 10% goat serum, 0.1% TX-100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample
Mouse Cell (primary embryonic epicardial)
Specification
primary embryonic epicardial
Permeabilization
Yes - 0.5% Triton X-100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 30 2014

1-10 of 26 Abreviews

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

Sign up