Phosphothreonine (incl. pos. control) Mouse Monoclonal Antibody [Clone ID: 4D11]

Specifications

Product Data
Clone Name	4D11
Applications	ELISA, IP, WB
Recommend Dilution	Immunoblotting (Western Blot): 1 µg/ml for HRPO/ECL detection. Recommended blocking buffer: BSA/Tween 20 based blocking buffer. DO NOT USE MILK OR CASEIN FOR BLOCKING! ELISA: 0.05 µg/ml. Immunoprecipitation: 1-10 µg per 10e6 pervanadate-treated A431 cells. Positive Control: Phosphoserine/Phosphothreonine.
Reactivity	Canine, Human, Mouse, Rat
Host	Mouse
Clonality	Monoclonal
Immunogen	Synthetic phosphopeptide conjugated to KLH.
Specificity	This antibody recognizes a broad range of threonine-phosphorylated proteins in crude cell extracts, providing a valuable tool for non-radioactive phosphothreonine detection.
Formulation	1 ml 2 x PBS / 0.09% Sodium Azide / PEG and Sucrose. State: Purified State: Lyophilized purified IgG fraction.
Reconstitution Method	Restore with 1 ml H2O (15 min, RT).
Purification	Size Exclusion Chromatography.
Conjugation	Unconjugated
Storage Condition	Store lyophilized (preferably in a desiccator) at -20°C and reconstituted (aliquote and freeze in liquid nitrogen) at -80°C. Avoid repeated freezing and thawing. Thaw aliquots at 37°C. Thawed aliquots may be stored at 4°C up to 1 week.
Background	Phosphorylation and dephosphorylation of cellular proteins are central steps in transducing extracellular signals to the cell nucleus. Phosphorylated epitopes may serve as docking sites for the assembly of protein complexes or may alter the 3-dimensional protein structure thus modulating enzymatic activity or the ability to undergo protein-protein-interactions. Modification of proteins on serine residues is mediated by serine/threonine kinases.
Note	This product contains a Positive Control (for details see "Protocols") Protocol: Positive Control: pSer / pThr Molecular Weight Marker Formulation: The pSer/pThr molecular weight marker contains rabbit muscle phosphoproteins isolated by Fe3+/IDA - affinity chromatography. Proteins are lyophilized from PBS/NaF/PEG/Sucrose/ Bromophenolblue and Na - azide. After reconstitution the solution contains 0.09% Na-azide. Stability: Reconstitute by addition of 200 µl H2O. After complete solubilization add 200 µl 2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Application: The pSer/pThr molecular weight marker is recommended for immunoblot applications. Use 20µl molecular weight marker per lane. Note: Use BSA based blot incubation buffers. Milk, Casein and Blotto might interfere with antibody - antigen interaction. Storage: Aliquote and store frozen. Avoid repeated freeze/thaw cycles. Shelf life: one year from despatch.
Reference Data

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