Phosphothreonine Antibody - Azide and BSA Free

Novus Biologicals | Catalog # NBP1-78035

Novus Biologicals

Key Product Details

Validated by

Biological Validation

Species Reactivity

Non-species specific

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

Azide and BSA Free
View all Phosphothreonine Primary Antibodies »

Product Specifications

Immunogen

Phosphothreonine Antibody was prepared from whole rabbit serum produced by repeated immunizations with phosphothreonine conjugated KLH.

Reactivity Notes

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

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This antibody was prepared from monospecific antiserum by immunoaffinity chromatography using phosphorylated threonine peptide coupled to agarose. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit IgG

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 - Azide and BSA Free

Western Blot: Phosphothreonine Antibody [NBP1-78035]

Western Blot: Phosphothreonine Antibody [NBP1-78035]

Western Blot: Phosphothreonine Antibody [NBP1-78035] - Goat anti Glucoamylase antibody was used to detect purified glucoamylase under reducing (R) and non-reducing (NR) conditions. Reduced samples of purified target proteins contained 4% BME and were boiled for 5 minutes. Samples of ~1ug of protein per lane.
Western Blot: Phosphothreonine Antibody [NBP1-78035]

Western Blot: Phosphothreonine Antibody [NBP1-78035]

Western Blot: Phosphothreonine Antibody [NBP1-78035] - shown to detect phosphorylated proteins present in mouse spleen cell lysates. Left Lane: extract of spleen from mouse treated with vanadium. Right lane: extract of spleen from untreated mouse.

Applications for Phosphothreonine Antibody - Azide and BSA Free

Application
Recommended Usage

ELISA

0.5 ug/ml

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

10.0 ug/mg

Western Blot

2.0 ug/ml
Application Notes
This affinity-purified antibody is tested for use in ELISA, western blotting and immunoprecipitation assays. The antibody reacts specifically with phosphothreonine and shows minimal reactivity by ELISA and western blotting with proteins containing phosphotyrosine or phosphoserine residues. The antibody reacts with free phosphothreonine, phosphothreonine conjugated to carriers such as thyroglobulin or BSA, and detects the presence of phosphothreonine in proteins of both unstimulated and stimulated cell lysates. Although not tested, this antibody is likely functional in RIA, flow cytometry and immunohistochemistry.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 50% (v/v) Glycerol

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

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 - Azide and BSA Free

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Product Specific Notices for Phosphothreonine Antibody - Azide and BSA Free

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.

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