Recombinant Anti-SH3PX1/SNX9 antibody [EPR14399] KO Tested (ab181856)

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Western blot - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR14399] to SH3PX1/SNX9
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Human

Carrier Free

Product name

Anti-SH3PX1/SNX9 antibody [EPR14399]
See all SH3PX1/SNX9 primary antibodies
Description

Rabbit monoclonal [EPR14399] to SH3PX1/SNX9
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Human
Predicted to work with: Rat
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- HeLa and HCT-116 cells and cell lysates; Human colon tissue.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR14399
Isotype

IgG
Research areas

Alternative Versions
- Anti-SH3PX1/SNX9 antibody [EPR14399] - BSA and Azide free (ab250527)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181856 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 (Intra)		1/10. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB		1/1000 - 1/10000. Detects a band of approximately 78 kDa (predicted molecular weight: 67 kDa).
IHC-P		1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/100.
IP		1/50.

Notes
Flow Cyt (Intra) 1/10. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/10000. Detects a band of approximately 78 kDa (predicted molecular weight: 67 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/50.

Function

May be involved in several stages of intracellular trafficking. Plays a role in endocytosis via clathrin-coated pits, but also clathrin-independent, actin-dependent fluid-phase endocytosis. Plays a role in macropinocytosis. Promotes internalization of TNFR. Promotes degradation of EGFR after EGF signaling. Stimulates the GTPase activity of DNM1. Promotes DNM1 oligomerization. Promotes activation of the Arp2/3 complex by WASL, and thereby plays a role in the reorganization of the F-actin cytoskeleton. Binds to membranes enriched in phosphatidylinositol 4,5-bisphosphate and promotes membrane tubulation. Has lower affinity for membranes enriched in phosphatidylinositol 3-phosphate.
Sequence similarities

Belongs to the sorting nexin family.
Contains 1 BAR domain.
Contains 1 PX (phox homology) domain.
Contains 1 SH3 domain.
Domain

The PX domain mediates interaction with membranes enriched in phosphatidylinositol phosphate. Has high affinity for phosphatidylinositol-4,5-bisphosphate, but can also bind to membranes enriched in other phosphatidylinositol phosphates.
Post-translational
modifications

Ubiquitinated by ITCH.
Phosphorylated on tyrosine residues by TNK2. Phosphorylation promotes its activity in the degradation of EGFR.
Cellular localization

Cytoplasmic vesicle membrane. Cell membrane. Cytoplasmic vesicle > clathrin-coated vesicle. Golgi apparatus > trans-Golgi network. Cell projection > ruffle. Cytoplasm. Localized at sites of endocytosis at the cell membrane. Detected on newly formed macropinosomes. Transiently recruited to clathrin-coated pits at a late stage of clathrin-coated vesicle formation. Colocalizes with the actin cytoskeleton at the cell membrane.
Target information above from: UniProt accession Q9Y5X1 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 51429 Human
- Entrez Gene: 66616 Mouse
- Entrez Gene: 683687 Rat
- Omim: 605952 Human
- SwissProt: Q9Y5X1 Human
- SwissProt: Q91VH2 Mouse
- Unigene: 191213 Human
- Unigene: 89515 Mouse
Alternative names
- MST 155 antibody
- MST155 antibody
- MSTP 155 antibody
- MSTP155 antibody
- OTTHUMP00000040083 antibody
- Protein SDP 1 antibody
- Protein SDP1 antibody
- SDP 1 antibody
- SDP1 antibody
- SH3 and PX domain containing protein 3A antibody
- SH3 and PX domain containing protein SH3PX1 antibody
- SH3 and PX domain-containing protein 1 antibody
- SH3 and PX domain-containing protein 3A antibody
- SH3PX1 antibody
- SH3PXD3A antibody
- SNX 9 antibody
- SNX9 antibody
- SNX9_HUMAN antibody
- Sorting nexin 9 antibody
- Sorting nexin-9 antibody
- WASP interactor protein antibody
- Wiskott Aldrich syndrome protein (WASP) interactor protein antibody
- Wiskott Aldrich syndrome protein interactor protein antibody
- WISP antibody
see all

Western blot - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: SH3PX1/SNX9 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HCT116 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab181856 observed at 75 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181856 was shown to specifically react with SH3PX1/SNX9 when SH3PX1/SNX9 knockout samples were used. Wild-type and SH3PX1/SNX9 knockout samples were subjected to SDS-PAGE. ab181856 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunoprecipitation - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Western blot analysis of HepG2 cell lysate immunoprecipitated using ab181856 at 1/50 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate secondary antibody was used at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Intracellular flow cytometric analysis of 2%paraformaldehyde-fixedHeLa cells labeling SH3PX1/SNX9 with ab181856 at 1/10 dilution (red) compared to a Rabbit IgG monoclonal isotype control (green), followed by Goat anti rabbit IgG (FITC) secondary antibodyat 1/150 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Immunofluorescent analysis of 4% paraformaldehyde-fixed HCT-116 cells labeling SH3PX1/SNX9 with ab181856 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®488) secondary antibody at 1/200 dilution. Counter stained with Dapi.
Immunocytochemistry/ Immunofluorescence - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Immunofluorescent analysis of acetone-fixed HeLa cells labeling SH3PX1/SNX9 with ab181856 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®488) secondary antibody at 1/200 dilution. Counter stained with Dapi.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SH3PX1/SNX9 with ab181856 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Western blot - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856) at 1/10000 dilution + HCT-116 cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 67 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?
Western blot - Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856) at 1/50000 dilution + HeLa cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 67 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?
Anti-SH3PX1/SNX9 antibody [EPR14399] (ab181856)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab181856? Please let us know so that we can cite the reference in this datasheet.

ab181856 has been referenced in 4 publications.

Ecker M et al. SNX9-induced membrane tubulation regulates CD28 cluster stability and signalling. Elife 11:N/A (2022). PubMed: 35050850
Jarsch IK et al. A direct role for SNX9 in the biogenesis of filopodia. J Cell Biol 219:N/A (2020). PubMed: 32328641
Shi J et al. Tanshinone IIA down-regulated p-Smad3 signaling to inhibit TGF-ß1-mediated fibroblast proliferation via lncRNA-HSRL/SNX9. Int J Biochem Cell Biol 129:105863 (2020). PubMed: 33049375
Mygind KJ et al. Sorting nexin 9 (SNX9) regulates levels of the transmembrane ADAM9 at the cell surface. J Biol Chem 293:8077-8088 (2018). PubMed: 29622675

Customer reviews and Q&As

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Positive Controls

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (4)

Customer reviews and Q&As

Post-translational
modifications