Thiophosphate Ester (WB version) antibody RabMAb®

Cat.#: 2686-1
Abcam ID: ab92570

Rabbit Monoclonal Antibody

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Description

This novel rabbit monoclonal antibody (RabMAb®) can be used to detect kinase substrates containing thiophosphate esters following the kinase reaction as described by J. Allen and colleagues in Nature methods 2007 (Figure 1). This antibody allows researchers to follow this approach to study kinase-substrate relationships of interest. The antibody is also context independent in that it can recognize serine, threonine, and tyrosine thiophosphorylated residues in a multitude of kinase phosphorylation consensus motifs (1).

This product is recommended for WB usage.

Recommended Applications

WB

Applications and Recommended Dilution Factors

WB: 1:1000 - 10,000

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Products Data

Figure 2. Western Blot analysis using Thiophosphate Ester Specific RabMAb (Cat# 2686-1)*. In vitro kinase reaction using JNK1 protein (10ng) incubated with c-jun-GST substrate (1ug) using ATP-gamma-S followed by alkylation with PNBM (2.5mM). Kinase reaction was carried out for 30 minutes at room temperature. PNBM alkylation (2.5mM) was for 2 hours at room temperature. Reaction products were analyzed by western blot with thiophosphate ester specific RabMAb (Cat# 2686-1) at 1:5000, ?-GST antibody confirms equal loading. Note: PNBM also reacts with cysteine residues and may decrease the affinity of other antibody-antigen interactions.

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Specificity

This antibody is specific to thiophosphorylated amino acids.

Storage Condition and Buffer

Store at -20 °C. Buffer: Antibody buffer, sodium azide, glycerol, and BSA. Stable for 12 months from date of receipt.

Description References

1. Allen, J.J. et al. Nature Methods, 4(6), 511-516, 2007.

Product References

1. Carlson SM, Chouinard CR, et al, Large-Scale Discovery of ERK2 Substrates Identifies ERK-Mediated Transcriptional Regulation by ETV3 ,Sci Signal. (2011) 4(196):rs11
2. Carlson HK, Plate L, Price MS, et al, Use of a semisynthetic epitope to probe histidine kinase activity and regulation ,Anal Biochem 397 (2010) 139-143
[Application: WB]
3. huluunbaatar U, Roller R, Feldman ME, et al, Constitutive mTORC1 activation by a herpesvirus Akt surrogate stimulates mRNA translation and viral replication ,Genes Dev 24 (2010) 2627-2639

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