概述

  • 产品名称Anti-SNX27抗体[1C6]
    参阅全部 SNX27 一抗
  • 描述
    小鼠单克隆抗体[1C6] to SNX27
  • 特异性Abcam scientists found that non-specific bands were eliminated by blocking with milk but not with BSA.
  • 经测试应用适用于: Flow Cyt, WB, IP, ICC/IFmore details
  • 种属反应性
    与反应: Human, Monkey
    不与反应: Mouse
  • 免疫原

    Fusion protein corresponding to Human SNX27 (N terminal).

  • 表位N-terminal amino acids 1-267.
  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; A431; HEK293; A549; MDA-MB-231.
  • 常规说明

    Alternative versions available:

    Anti-SNX27 antibody [1C6] - BSA and Azide free (ab173233)

    Anti-SNX27 antibody (HRP) [1C6] (ab203648)

性能

应用

Our Abpromise guarantee covers the use of ab77799 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
Flow Cyt Use 0.1-1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 10 µg/ml. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).

Abcam recommends using 3% milk as the blocking agent.

IP Use at an assay dependent concentration.
ICC/IF 1/500. Use with paraformaldehyde- or methanol- fixed cells.

靶标

  • 功能Involved in endocytic trafficking (By similarity). In T lymphocytes, participates in endocytic recycling pathway. Recruits PSCDBP and HT4R to early endosomes.
  • 组织特异性Expressed in cells of hematopoietic origin (at protein level).
  • 序列相似性Belongs to the sorting nexin family.
    Contains 1 PDZ (DHR) domain.
    Contains 1 PX (phox homology) domain.
    Contains 1 Ras-associating domain.
  • 结构域The PDZ domain mediates the interaction with DGKZ, PSCDBP and HT4R and is responsible for vesicular localization.
  • 细胞定位Cytoplasm > cytosol. Early endosome. In T-lymphocytes, recruited from the cytosol to sorting endosomes by phosphoinositide-3-kinase products.
  • Information by UniProt
  • 数据库链接
  • 别名
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    • SNX27 antibody
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Anti-SNX27 antibody [1C6] 图像

  • All lanes : Anti-SNX27 antibody [1C6] (ab77799) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
    Lane 5 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 61 kDa
    Observed band size : 61 kDa


    Exposure time : 15 minutes
  • ab77799 (1/500) detecting SNX27 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab77799 (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 (ab77799, 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 Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • SNX27 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to SNX27 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab77799.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 61kDa: SNX27.

Anti-SNX27 antibody [1C6] (ab77799)参考文献

This product has been referenced in:
  • Singh V  et al. Sorting nexin 27 regulates basal and stimulated brush border trafficking of NHE3. Mol Biol Cell 26:2030-43 (2015). WB . Read more (PubMed: 25851603) »
  • Bauch C  et al. Subcellular sorting of the G-protein coupled mouse somatostatin receptor 5 by a network of PDZ-domain containing proteins. PLoS One 9:e88529 (2014). WB . Read more (PubMed: 24523912) »

See all 6 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions Reduced Denaturing
Loading amount 25 µg
Specification HeLa cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 17 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 20°C
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.1% Triton TX-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 May 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293T cells)
Loading amount 25 µg
Specification HEK293T cells
Gel Running Conditions Reduced Denaturing (4-12% SDS-PAGE gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Jan 24 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (brain and PC12 cells)
Loading amount 40 µg
Specification brain and PC12 cells
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Mar 30 2012

Thank you very much for your interest in our SNX27 antibodies.

To our knowledge, neither ab77799 nor ab103354 have been tested in rat yet, but that does not mean they might not work.

I absolutely agree that with an immunogen seque...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100
Username

Dr. Kirk McManus

Verified customer

提交于 Feb 01 2011

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