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P6623

Sigma-Aldrich

Anti-Phosphothreonine antibody, Mouse monoclonal

clone PTR-8, purified from hybridoma cell culture

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Synonym(s):
Monoclonal Anti-Phosphothreonine, Phospho Thr, Phospho Threonine, Phospho-Thr, Phospho-Threonine, p-Thr

biological source

mouse

Quality Level

conjugate

unconjugated

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

PTR-8, monoclonal

form

buffered aqueous solution

concentration

~2.5 mg/mL

technique(s)

dot blot: suitable
immunocytochemistry: suitable
immunoprecipitation (IP): suitable
indirect ELISA: 0.5-1 μg/mL
microarray: suitable
western blot: 5-10 μg/mL using A431 cell extracts

isotype

IgG2b

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

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This Item
P5872P1869P5747
antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

clone

PTR-8, monoclonal

clone

PT-66, monoclonal

clone

pT-154, monoclonal

clone

PSR-45, monoclonal

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

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General description

As determined by ELISA and dot blot, the antibody reacts specifically with phosphorylated threonine, both as free amino acid or conjugated to carriers such as BSA or KLH. No cross-reactivity is observed with non-phosphorylated threonine, phosphoserine, phosphotyrosine, AMP or ATP. This antibody has been used in immunoblotting for the localization of some phosphothreonine-containing proteins. Certain proteins known to contain phosphorylated threonine may not be recognized by this antibody due to steric hindrance of the recognition site.
Monoclonal Anti-Phosphothreonine (mouse IgG2b isotype) is derived from the hybridoma produced by the fusion of mouse myeloma cells (NS1) and splenocytes from BALB/c mice immunized with phosphothreonine conjugated to keyhole limpet hemocyanin (KLH).
Mouse monoclonal clone PTR-8 anti-Phosphothreonine antibody reacts with phosphorylated threonine both as a free amino acid or when conjugated to carriers such as BSA or KLH, using ELISA and dot blot. It does not react with nonphosphorylated threonine, phosphorylated tyrosine or serine, AMP or ATP.

Specificity

Monoclonal Anti-Phosphothreonine reacts with phosphorylated threonine both as a free amino acid or when conjugated to carriers such as BSA or KLH, using ELISA and dot blot. It does not react with nonphosphorylated threonine, phosphorylated tyrosine or serine, AMP or ATP.

Immunogen

phosphothreonine conjugated to keyhole limpet hemocyanin (KLH).

Application

Monoclonal Anti-Phosphothreonine antibody produced in mouse has been used in enzyme-linked immunosorbent assay (ELISA), dot blot, immunoprecipitation, immunocytochemistry and immunoblotting. It has also been used in Akt kinase assay to detect phosphorylation status on serine and threonine residue in the wild type or mutant type δ-catenin.

Biochem/physiol Actions

Protein phosphorylation and dephosphorylation are basic mechanisms for the modification of protein function in eukaryotic cells. Phosphorylation is a rare post-translational event in normal tissue. Various activation processes are mediated through phosphotyrosine, phosphoserine or phosphothreonine (p-Tyr/p-Ser/p-Thr). Many different mitogenic systems, such as the epidermal growth factor (EGF), platelet derived growth factor (PDGF) and insulin receptor systems contain Tyr/Ser/Thr kinase domains that autophosphorylate specific Tyr/Ser/Thr residues upon binding of their ligands. T cell antigen receptor complex or receptors for some hemopoietic growth factors may stimulate associated kinases, and cells transformed by viral oncogenes contain elevated levels of phosphorylated Tyr/Ser/Thr.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Storage and Stability

For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze at -20 °C in working aliquots. Repeated freezing and thawing, or storage in "frost-free" freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

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.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable


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Kim H, et al.
The Journal of Biological Chemistry, 283(2), 977-987 (2008)
The origins of protein phosphorylation
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Nature Cell Biology, 4(5), E127-E127 (2002)

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