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Phosphothreonine Antibody (18F6)

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Western Blot: Phosphothreonine Antibody (18F6) [NB600-878] - Phospho Threonine monoclonal antibody, clone 18F6 is shown to detect threonine phosphorylation of proteins in a lysate from A-431 cells stimulated with EGF.
ELISA: Phosphothreonine Antibody (18F6) [NB600-878] - was tested against BSA conjugates of phosphothreonine (grey squares), phosphotyrosine (black triangles) and phosphoserine (open circles). Each well was coated with ...read more
Mab Anti-phosphothreonine Antibody(clone 18F6) is shown to detect threonine phosphorylation of proteins in a lysate from A431 cells stimulated with EGF

Product Details

Summary
Reactivity All-NASpecies Glossary
Applications WB, ELISA
Clone
18F6
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

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Phosphothreonine Antibody (18F6) Summary

Description
Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
Immunogen
Phosphothreonine Antibody (18F6) was produced after repeated immunizations of balb/c mice with phosphothreonine conjugated KLH.
Specificity
Reactivity is specific for phosphothreonine and minimal cross reactivity is observed against phosphoserine or phosphotyrosine.
Isotype
IgG1 Kappa
Clonality
Monoclonal
Host
Mouse
Purity
Unpurified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • ELISA 1:4000-1:20000
  • Western Blot 1:500-1:2000
Application Notes
This monoclonal antibody has been tested in ELISA and Western Blot and reacts specifically with phosphothreonine and shows minimal reactivity by ELISA, and competitive ELISA, with phosphoserine or phosphotyrosine. The antibody reacts with free phosphothreonine, phosphothreonine conjugated to carriers such as thyroglobulin or BSA, as well as phosphothreonine in proteins of both unstimulated and stimulated cell lysates. Although not tested, this antibody is likely functional in RIA, flow cytometry, immunohistochemistry and immunoprecipitation. Phosphorylation of threonine residues is associated with many growth factors and oncogene protein kinases and is important for cell signaling in activation, proliferation and differentiation.
Publications
Read Publication using NB600-878.

Reactivity Notes

Reactivity is specific for phosphothreonine and minimal cross reactivity is observed against phosphoserine or phosphotyrosine

Packaging, Storage & Formulations

Storage
Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
Buffer
Antiserum
Preservative
0.01% Sodium Azide
Purity
Unpurified

Alternate Names for Phosphothreonine Antibody (18F6)

  • C4H10NO6P;O-phospho-L-threonine;Phospho-threonine;pThr;Threonine

Background

Phosphorylation of threonine residues is associated with many growth factors and oncogene protein kinases, and is important for cell signaling in activation, proliferation and differentiation. 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, however, the abundance of phosphorylated cellular proteins increases several fold following various activation processes which are mediated through phosphotyrosine, phosphoserine or phosphothreonine (p-tyr/p-ser/p-thr). Many signal transduction pathways, such as the EGF, PDGF and insulin receptor systems, contain tyr/ser/thr kinase which phosphorylate specific tyr/ser/thr residues upon binding of ligands to their receptors. T cell antigen receptor complex or the receptors for some hemopoietic growth factors may stimulate these phosphorylation associated kinases, and cells transformed by viral oncogenes contain elevated levels of phosphorylated tyr/ser/thr. An understanding of transformation by oncogenes and mitogenic processes of growth factors depends on the identification of their substrate and a subsequent determination of how phosphorylation affects their properties. Studies on the role of phosphorylated proteins have been hampered by their low abundance and the problem of distinguishing the various types of phosphorylated proteins. The most common procedure is to label intact cells or small tissue fragments with 32P and subsequently to isolate 32P labeled proteins by conventional biochemical methods. In order to identify the specific amino acids that undergo phosphorylation, additional long and tedious procedures for phosphoamino acid analysis are required. Immunoblotting of cellular proteins with antibodies directed against phosphoamino acids is advantageous as it does not involve 32P labeling, and can therefore be employed to monitor alterations in phosphorylation of specific proteins as they occur in intact organs or the whole animal. Indeed, mono and polyclonal antibodies directed against phosphorylated residues have been generated and found useful as analytical and preparative tools because they enable the rapid identification, quantification and immunoaffinity isolation of phosphorylated cellular proteins.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Phosphothreonine Antibody (NB600-878)(1)

We have publications tested in 1 application: WB.


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Product General Protocols

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Video Protocols

WB Video Protocol

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Secondary Antibodies

 

Isotype Controls

Additional Phosphothreonine Products

Array NB600-878

Research Areas for Phosphothreonine Antibody (NB600-878)

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Blogs on Phosphothreonine.

Phosphotyrosine: A Global Player in Human Health and Disease
Protein tyrosine phosphorylation is a fundamental mechanism for controlling many aspects of cellular processes, as well as aspects of human health and diseases. Compared with phosphoserine and phosphothreonine, phosphotyrosine signaling is more tightl...  Read full blog post.

Phosphotyrosine: Affecting Locally Acting Globally
Over the past two decades, it has become clear that tyrosine phosphorylation plays a pivotal role in a variety of important signaling pathways in multicellular organisms. In the typical vertebrate cell, phosphotyrosine represents only a tiny fraction ...  Read full blog post.

Phosphoserine: A Basic Cellular Signal with Wide Ranging Effects
Phosphoserine is an ester of serine and phosphoric acid which results from posttranslational modifications. Phosphorylation is a key post-translational modification necessary for normal cellular signaling. Phosphorylated proteins mediate cell division...  Read full blog post.

Phosphotyrosine is Critical Signal Transduction and Regulation
Phosphotyrosine is the phosphorylated version of the amino acid tyrosine, which results from the activation of intracellular protein kinases (e.g. via growth factors) during normal growth and development, well as in transformation and oncogenesis. Pho...  Read full blog post.

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