Product nameAnti-Phosphoserine/threonine/tyrosine antibody [SPM101]
DescriptionMouse 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 reactivityReacts with: Species independent
Phosphoserine, phosphothreonine, and phosphotyrosine conjugated to protein carriers.
- Breast carcinoma
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Concentration information loading...
PurityProtein A/G purified
Purification notesPurified from ascites fluid by Protein G.
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.
|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|
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.
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.
This product has been referenced in:
- Pereira C et al. Sit4p-mediated dephosphorylation of Atp2p regulates ATP synthase activity and mitochondrial function. Biochim Biophys Acta 1859:591-601 (2018). Read more (PubMed: 29719209) »
- Vinyoles M et al. Activation of CK1? by PP2A/PR61? is required for the initiation of Wnt signaling. Oncogene 36:429-438 (2017). Human . Read more (PubMed: 27321178) »