Anti-Alpha SNAP抗体[15D4] (ab28352)

概述

  • 产品名称Anti-Alpha SNAP抗体[15D4]
    参阅全部 Alpha SNAP 一抗
  • 描述
    小鼠单克隆抗体[15D4] to Alpha SNAP
  • 特异性ab28352 specifically recognizes alpha-SNAP as a single band of ~36kDa on western blot and does not recognize beta-SNAP.
  • 经测试应用适用于: IHC-P, WB, IP, ICC/IF, Flow Cyt, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human
    不与反应: Chicken
  • 免疫原

    N-terminal domain between amino acids 1-109 of alpha-SNAP

  • 阳性对照
    • Rat kidney, rat brain and MDBK cells. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal prostate.

性能

应用

Our Abpromise guarantee covers the use of ab28352 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
IHC-P Use a concentration of 5 µg/ml.
WB 1/5000. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa).
IP Use a concentration of 2 - 10 µg/ml.
ICC/IF 1/100.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC Use at an assay dependent concentration.

靶标

Anti-Alpha SNAP antibody [15D4] 图像

  • All lanes : Anti-Alpha SNAP antibody [15D4] (ab28352) at 0.5 µg/ml

    Lane 1 : Rat brain lysate
    Lane 2 : Rat kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP conjugated goat anti mouse IgG at 0.2 µg/ml

    Predicted band size : 33 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds
  • ab28352 staining alpha SNAP in cultured Human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X (0.5%) and blocked with 3% BSA for 1 hour at 24°C. Samples were incubated with primary antibody (1/100) for 1 hour at 24°C. An Alexa Fluor®488-conjugated Chicken anti-mouse polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab28352 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 followed by the antibody (ab28352, 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 IgG1 [ICIGG1] (ab91353, 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 with 0.1% PBS-Tween for 20 min used under the same conditions.

  • IHC image of Alpha SNAP staining in Human normal prostate formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28352, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-Alpha SNAP antibody [15D4] (ab28352)参考文献

ab28352 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - other (MEF)
Loading amount 30 µg
Specification MEF
Gel Running Conditions Reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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提交于 Nov 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Mouse Cultured Cells (MEF)
Specification MEF
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton-X
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
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提交于 Oct 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - other (HELA)
Total protein in input 200 µg
Specification HELA
Immuno-precipitation step Protein G
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提交于 Oct 05 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cultured Cells (HELA)
Specification HELA
Fixative Paraformaldehyde
Permeabilization Yes - Triton-X-0.5%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Oct 05 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Jurkat T lymphocytes)
Loading amount 50 µg
Specification Jurkat T lymphocytes
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
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Verified customer

提交于 Oct 05 2010

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