Phosphothreonine Antibody (18F6)

Novus Biologicals | Catalog # NB600-878

Novus Biologicals
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Key Product Details

Species Reactivity

Non-species specific

Applications

Validated:

Western Blot, ELISA

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 18F6
View all Phosphothreonine Primary Antibodies »

Product Specifications

Immunogen

Phosphothreonine Antibody (18F6) was produced after repeated immunizations of balb/c mice with phosphothreonine conjugated KLH.

Reactivity Notes

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

Specificity

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

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

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.

Scientific Data Images for Phosphothreonine Antibody (18F6)

Western Blot: Phosphothreonine Antibody (18F6) [NB600-878]

Western Blot: Phosphothreonine Antibody (18F6) [NB600-878]

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]

ELISA: Phosphothreonine Antibody (18F6) [NB600-878]

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 0.1 ug of conjugate. The starting dilution of antibody was 1/1000 and each point on the X axis represents a 2-fold dilution. For detection, an HRP-conjugated Goat anti-Mouse IgG was used as secondary antibody, with a TMB substrate. Absorbance is represented on the Y axis.
Phosphothreonine Antibody (18F6)

Phosphothreonine Antibody (18F6)

Mab Anti-phosphothreonine Antibody(clone 18F6) is shown to detect threonine phosphorylation of proteins in a lysate from A431 cells stimulated with EGF

Applications for Phosphothreonine Antibody (18F6)

Application
Recommended Usage

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.

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Antiserum

Preservative

0.01% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: Phosphothreonine

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.

Alternate Names

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

Additional Phosphothreonine Products

Product Documents for Phosphothreonine Antibody (18F6)

Certificate of Analysis

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Product Specific Notices for Phosphothreonine Antibody (18F6)

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.

Citations for Phosphothreonine Antibody (18F6)

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