Analysis Note

ControlNIH 3T3 cells (+/- TPA), K562 cells, and EGF-stimulated A431 cells.

Application

Research CategorySignaling

Research Sub CategoryGeneral Post-translation Modification

Immunoprecipitation (tissue extracts):
10-20 µg anti-pS/mg protein was used from a previous lot.
Note that this will immunoprecipitate all phosphoserine proteins in the extracts. If using purified protein exacts containing only the protein of interest, antibody amounts should be decreased to 5 µg/500 µL of extracts. Optimal concentrations must be determined experimentally.

ELISA (kinase assay):
A previous lot of this anitbody was used at a 1:250-1:500 dilution.

Immunohistochemistry:
A previous lot of this antibody was used at a 1:50 dilution.

As the antibody detects all phosphoserines, any phosphorylated serine protein or peptide can be used to block antibody staining.
Typically a 10 M excess of peptide is used in Tris based buffers.

Western Blot Analysis:
1:500 dilution of a previous lot

Optimal working dilutions must be determined by end user.

Anti-Phosphoserine Antibody detects level of Phosphoserine & has been published & validated for use in ELISA, IH, IP & WB.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Cellular responses to a variety of extracellular signals occur through phosphorylation or dephosphorylation of intracellular proteins. Abnormal protein phosphorylation may be involved in the progression of numerous diseases, including many forms of cancer, immune system dysfunction and cardiovascular disease. Such changes in protein phosphorylation may be detected by immunocytochemical mapping of cells labeled with anti-phosphoserine or anti-phosphothreonine antibodies and can be correlated with cellular, electrophysiological or behavioral state changes.

Immunogen

Keyhole Limpet Hemocyanin(KLH)- conjugated phosphoserine conjugates.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified rabbit serum in buffer containing 50% glycerol.

Phosphoserine affinity chromatography

Quality

Routinely evaluated by Western Blot on NGF treated PC12 lysates.

Western Blot Analysis: 1:500 dilution of this lot detected serine phosphorylated proteins on 10 µg of NGF treated PC12 lysates.

Specificity

Anti-phosphoserine is species independent.

Recognizes phosphoserine, peptidyl-phosphoserine, and serine-phosphorylated proteins. Does not cross react with ATP, phosphotyrosine, peptidyl phosphothreonine and serine. Slight cross reactivity with free phosphothreonine. Readily reacts with known phosphoproteins such as phosvitin and alpha casein. Anti-phosphoserine is species independent in reactivity

Storage and Stability

Stable for up to 6 months at -20°C in undiluted aliquots from date of receipt. During shipment, small volumes of product will occasionally become entrapped in the seal of the product vial. For products with volumes of 200 uL or less, we recommend gently tapping the vial on a hard surface or briefly centrifuging the vial in a tabletop centrifuge to dislodge any liquid in the container′s cap.

Target description

Dependent upon the molecular weight of the serine phosphorylated protein being detected.