概述

  • 产品名称
    Anti-SM22 alpha抗体
    参阅全部 SM22 alpha 一抗
  • 描述
    兔多克隆抗体to SM22 alpha
  • 宿主
    Rabbit
  • 特异性
    Recognises a band at 23kD that is specifically blocked by the immunising peptide.
  • 经测试应用
    适用于: ICC/IF, IHC-P, IHC-Fr, WB, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Cow, Human, Pig
  • 免疫原

    Synthetic peptide, conjugated to KLH derived from within residues 150 - 200 of Mouse SM22 alpha.

    .

  • 阳性对照
    • This antibody gave a positive signal the following lysates for Western Blot: HeLa; HeLa Nuclear; Human Skeletal Muscle; Human Colon; Mouse Colon; Rat Colon ICC/IF: Hela cells

性能

应用

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.

应用 Ab评论 说明
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.

靶标

图片

  • ab14106 stained Hela cells. The cells were 100% methanol fixed for 5 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 min 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 1µg/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 2/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour 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.

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  • All lanes : Anti-SM22 alpha antibody (ab14106) at 2 µg/ml

    Lane 1 : HeLa 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



    Lane 1 - 3 : SM22 alpha antibody (ab14106) at 1 ug/ml

    Lane 1 : HeLa Whole Cell Lysate at 20 ug
    Lane 2 : HeLa Nuclear Extract at 20 ug
    Lane 3 : Human Muscle lysate at 20 ug

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

    Observed band size : 23kD

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

     


     

     

     

  • ab14106, at 1/100, staining SM22 alpha in chicken smooth muscle tissue sections by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were methacarn (60% MeOH, 30% choloform, 10% acetic acid) fixed, prior to blocking in 2% serum for 5 minutes at 25°C and then incubated with ab14106, for 1 hour at 25°C. Alexa fluor® 488 goat polyclonal to rabbit Ig, diluted 1/100, was used as the secondary antibody.

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

    All lanes : Lysates prepared from primary Human 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

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

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

文献

This product has been referenced in:

See all 131 Publications for this product

客户评价及客户问答

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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
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提交于 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
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提交于 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
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提交于 Mar 15 2018

Application
IHC - Wholemount
Sample
Mouse Tissue (artery)
Specification
artery
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提交于 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
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提交于 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
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提交于 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
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提交于 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
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提交于 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
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提交于 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
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提交于 Jan 30 2014

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