Anti-Phosphoserine/threonine/tyrosine antibody [SPM101] (ab15556)

Overview

  • Product nameAnti-Phosphoserine/threonine/tyrosine antibody [SPM101]
  • Description
    Mouse monoclonal [SPM101] to Phosphoserine/threonine/tyrosine
  • Tested applicationsSuitable for: IHC - Wholemount, Functional Studies, ICC/IF, Flow Cyt, ELISA, WB, IP, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Phosphoserine, phosphothreonine, and phosphotyrosine conjugated to protein carriers.

  • Positive control
    • Breast carcinoma

Properties

Applications

Our Abpromise guarantee covers the use of ab15556 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
IHC - Wholemount 1/200.
Functional Studies Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ELISA Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Milk derived blocking solutions reportedly contain phosphoproteins that may inhibit phosphoamino acid antibody binding and therefore should be avoided.
IP Use at 2 µg/mg of lysate.
IHC-P 1/20. Boil tissue sections in 10mM citrate, pH 6.0 for 10 min followed by cooling at RT for 20 min.
IHC-Fr Use at an assay dependent concentration. PubMed: 20421451

Target

  • RelevanceProtein phosphorylation provides a signalling system that can be thought of as a kind of protein on/off switch for many cellular signalling pathways. Phosphorylation is observed on serine, threonine, tyrosine and histidine residues. Cellular networks underlying phosphorylation can be very complex and often occurs on multiple distinct sites on a given protein. Phospho-specific antibodies are becoming critical reagents both for basic research and for clinical diagnosis.

Anti-Phosphoserine/threonine/tyrosine antibody [SPM101] images

  • Human breast carcinoma stained with ab15556. Immunohistochemistry (FFPE-sections).
  • Overlay histogram showing MCF7 cells stained with ab15556 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab15556, 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 MCF7 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Immunohistochemical analysis of mouse cardiomyocytes, staining Phosphoserine/threonine/tyrosine with ab15556.

    Samples were incubated with primary antibody (1/200 in diluent) for 1 hour. An AlexaFluor®488-conjugated goat anti-mouse polyclonal IgG (1/400) was used as the secondary antibody.

    See Abreview

References for Anti-Phosphoserine/threonine/tyrosine antibody [SPM101] (ab15556)

This product has been referenced in:
  • Ekyalongo RC  et al. TYRO3 as a potential therapeutic target in breast cancer. Anticancer Res 34:3337-45 (2014). Read more (PubMed: 24982338) »
  • Rocchetti MT  et al. Two dimensional gel phosphoproteome of peripheral blood mononuclear cells: comparison between two enrichment methods. Proteome Sci 12:46 (2014). WB ; Human . Read more (PubMed: 25276096) »

See all 8 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (colon cell)
Permeabilization Yes - 1% triton x100
Specification colon cell
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative Paraformaldehyde
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Verified customer

Submitted Sep 05 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (293T)
Gel Running Conditions Non-reduced Non-Denaturing (Native)
Loading amount 80 µg
Treatment damage stress
Specification 293T
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Sep 05 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (mice pancreas tumors)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
Permeabilization Yes - Tween-20
Specification mice pancreas tumors
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Formaldehyde
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Submitted Aug 18 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Human fibromatosis)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
Permeabilization Yes - Tween-20
Specification Human fibromatosis
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Formaldehyde
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Submitted Aug 18 2015

Thank you for contacting us.

The ab9337 recognizes proteins phosphorylated on threonine residues. It does not cross-react with phosphotyrosine.

For ab15556, this antibody recognizes serine, threonine, and tyrosine phosphorylated pr...

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It (ab15556) is specific for phosphate groups on any serine, threonine, or tyrosine, on any protein, so it may not be specific enough for your purposes, if your samples contain other phosphorylated proteins besides EGFR.

Thank you for contacting us.

I have been in touch with the originator of this product to discuss exactly how tab15556 Anti-Phosphoserine/threonine/tyrosine antibody [SPM101] is made. Apparently the immunogen was a carrier protein (used to eli...

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I am very pleased to hear you would like to accept our offer and test ab15556 in plants in WB. This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview forgrape extracta...

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Thank you for your reply.

Unfortunately, 2D gel analysis is not an application we can issue the Abreview discount for. The reason for this is that this is not an application you can review via our system.

However, I would gladly is...

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Thank you very much for your interest in ab15556 Anti-Phosphoserine/threonine/tyrosine antibody [SPM101].

To our knowledge, ab15556has not been tested on plants, and unfortunately, we do not have any reference for this. May I ask what specie...

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