Anti-Phosphothreonine Antibody

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.

Keyhole Limpet Hemocyanin (KLH) – conjugated phosphothreonine conjugates and Phosvitin.

Anti-Phosphothreonine Antibody is an antibody against Phosphothreonine for use in ELISA, IH(P), IP & WB.

ELISA (kinase assay):
A 0.5 μg/mL concentration of a previous lot was used in ELISA.

Immunoprecipitation:
A previous lot of this antibody was used in IP.

Immunohistochemistry (paraffin):
Representative image from a previous lot.

Optimal working dilutions must be determined by end user.

Research Category
Signaling

Research Sub Category
General Post-translation Modification

Anti-phosphothreonine is species independent

By direct competitive ELISA methodology (antigen capture) the specificity of this antibody has been determined to be phosphothreonine. Binding of Ly-pT-Gly-HRP (0.2μg/100μL/well) to an anti-phosphothreonine coated microplate (0.5μg/100μL/well) is inhibited by 95% with the addition of 2-10 μg/mL anti-phosphothreonine. No inhibition is evident by inhibition of phosphoserine, phosphotyrosine, or threonine at concentrations of up to 1 mg/mL. Antibody AB1607 recognizes phosphothreonine, peptidyl-phosphothreonine, and threonine-phosphorylated proteins.

ImmunoAffinity Purified

Purified rabbit polyclonal in PBS buffer, containing 50% Glycerol, 0.01% NaN3.

Stable for 1 year at -20ºC in undiluted aliquots from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Control
Mouse brain lysate.

Routinely evaluated by Western Blot on Calyculin/Okadaic Acid treated HEK293 lysate.

Western Blot Analysis: 1:500 dilution of this lot detected PHOSPHOTHREONINE on 10 μg of Calyculin/Okadaic Acid treated HEK293 lysate.

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

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

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.