Product nameAnti-SNX27 antibody [1C6]
See all SNX27 primary antibodies
DescriptionMouse monoclonal [1C6] to SNX27
SpecificityAbcam scientists found that non-specific bands were eliminated by blocking with milk but not with BSA.
Tested applicationsSuitable for: Flow Cyt, WB, IP, ICC/IFmore details
Species reactivityReacts with: Human, Monkey
Does not react with: Mouse
Fusion protein corresponding to Human SNX27 aa 1-300 (N terminal).
EpitopeN-terminal amino acids 1-267.
- This antibody gave a positive signal in the following whole cell lysates: HeLa; A431; HEK293; A549; MDA-MB-231.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
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.
|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.|
FunctionInvolved in endocytic trafficking (By similarity). In T lymphocytes, participates in endocytic recycling pathway. Recruits PSCDBP and HT4R to early endosomes.
Tissue specificityExpressed in cells of hematopoietic origin (at protein level).
Sequence similaritiesBelongs to the sorting nexin family.
Contains 1 PDZ (DHR) domain.
Contains 1 PX (phox homology) domain.
Contains 1 Ras-associating domain.
DomainThe PDZ domain mediates the interaction with DGKZ, PSCDBP and HT4R and is responsible for vesicular localization.
Cellular localizationCytoplasm > cytosol. Early endosome. In T-lymphocytes, recruited from the cytosol to sorting endosomes by phosphoinositide-3-kinase products.
- Information by UniProt
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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.
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.
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.
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) »