Overview

  • Product name

    Anti-SNX27 antibody [1C6]
    See all SNX27 primary antibodies
  • Description

    Mouse monoclonal [1C6] to SNX27
  • Host species

    Mouse
  • Specificity

    Abcam scientists found that non-specific bands were eliminated by blocking with milk but not with BSA.
  • Tested applications

    Suitable for: Flow Cyt, WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Human, Monkey
    Does not react with: Mouse
  • Immunogen

    Fusion protein corresponding to Human SNX27 aa 1-300 (N terminal).

  • Epitope

    N-terminal amino acids 1-267.
  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: HeLa; A431; HEK293; A549; MDA-MB-231.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function

    Involved in endocytic trafficking (By similarity). In T lymphocytes, participates in endocytic recycling pathway. Recruits PSCDBP and HT4R to early endosomes.
  • Tissue specificity

    Expressed in cells of hematopoietic origin (at protein level).
  • Sequence similarities

    Belongs to the sorting nexin family.
    Contains 1 PDZ (DHR) domain.
    Contains 1 PX (phox homology) domain.
    Contains 1 Ras-associating domain.
  • Domain

    The PDZ domain mediates the interaction with DGKZ, PSCDBP and HT4R and is responsible for vesicular localization.
  • Cellular localization

    Cytoplasm > cytosol. Early endosome. In T-lymphocytes, recruited from the cytosol to sorting endosomes by phosphoinositide-3-kinase products.
  • Information by UniProt
  • Database links

  • Alternative names

    • KIAA0488 antibody
    • Methamphetamine responsive transcript 1 antibody
    • MGC126871 antibody
    • MGC126873 antibody
    • MGC20471 antibody
    • MRT1 antibody
    • MY014 antibody
    • OTTHUMP00000042487 antibody
    • RP11-98D18.12 antibody
    • SNX27 antibody
    • SNX27_HUMAN antibody
    • Sorting nexin family member 27 antibody
    • Sorting nexin-27 antibody
    see all

Images

  • 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
    All lanes : 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


    Blocking: 5% Milk in TBS-T.

    Two bands are observed very close together, at around 60-kDa. There are four reported isoforms of the SNX27 protein (Swissprot). We hypothesise that this antibody detects both isoform 1 and isoform 2, which are 61-kDa and 60-kDa respectively. As the immunogen is derived from the N-terminal region of SNX27, the antibody is not expected to react with isoforms 3 or 4.

  • 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.

References

This product has been referenced in:

  • Binda CS  et al. Sorting nexin 27 rescues neuroligin 2 from lysosomal degradation to control inhibitory synapse number. Biochem J 476:293-306 (2019). Read more (PubMed: 30602588) »
  • Strutt H  et al. Retromer Controls Planar Polarity Protein Levels and Asymmetric Localization at Intercellular Junctions. Curr Biol 29:484-491.e6 (2019). Read more (PubMed: 30661800) »
See all 20 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Oct 17 2016

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

Abcam user community

Verified customer

Submitted May 23 2013

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

Abcam user community

Verified customer

Submitted Jan 24 2013

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

Abcam user community

Verified customer

Submitted Mar 30 2012

Answer

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 sequence similarity of 93% (i.e. 28 out of 30 amino acids) the antibody ab103354 may indeed cross-react with rat. Unfortunately, because we carry over 80,000 products, it isn't feasible for us to keep small sample sizes of our products. But I am happy to let you know that we have our Abcam guarantee in place (www.abcam.com/Abpromise), and for cases such as yours our testing discount programme:

As this antibody has not been tested in rat but might work, we can offer a discount off a future purchase if you buy the antibody now, test it in rat and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab103354 in rat. I will then send a discount code. This code must be issued before purchasing the antibody so please wait for my reply before ordering.

2. Purchase ab103354 either by phone, fax, or online (www.abcam.com).

3. Test it in rat.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews .

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab103354 turns out to be unsuitable for rat, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about these offers and I would be happy to help you further.

The Terms and Conditions can be found at: www.abcam.com/collaborationdiscount

PS: If you are happy with the results of ab77799, we would be very grateful if you could submit an Abreview about this product via the online product datasheet. You could earn quite a few Abpoints with it ;-)

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 01 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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