Overview

  • Product name
    Anti-Thiophosphate ester antibody [51-8]
    See all Thiophosphate ester primary antibodies
  • Description
    Rabbit monoclonal [51-8] to Thiophosphate ester
  • Host species
    Rabbit
  • Specificity
    This antibody is specific to thiophosphorylated amino acids.
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    not applicable
  • Immunogen

    Chemical/ Small Molecule corresponding to Thiophosphate ester conjugated to keyhole limpet haemocyanin.

  • General notes

    This antibody was developed as part of a collaboration between Abcam and the lab of Dr. Kevan Shokat at University of California at San Francisco.

    It is possible to use ab92570 with ab138911 as a kinase reaction reagent for the identification of direct kinase substrates as mentioned in Allen JJ et al. Nat Methods 4:511-6 (2007).

    After the kinase of interest has accepted ATP-γ-S (ab138911), p-Nitrobenzyl mesylate (ab138910) can be used to alkylate the thiophospholyation site on the substrates. ab92570 is introduced to identify the tagged substrates.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.5% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Primary antibody notes
    ab92570 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. This antibody as allow 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.
    In order for this antibody to work in WB and IP, the customer needs to perform a specific kinase reaction using ATP gamma S and PNBM.
    See specific Protocol at the bottom of this page for details.
  • Clonality
    Monoclonal
  • Clone number
    51-8
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab92570 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/5000.

Target

  • Relevance
    Thiophosphate ester is used as fungicide.

Images

  • All lanes : Anti-Thiophosphate ester antibody [51-8] (ab92570) at 1/5000 dilution

    Lane 1 : JNK1 protein (10ng) incubated with c-jun-GST substrate (1ug)
    Lane 2 : JNK1 protein (10ng) incubated with c-jun-GST substrate (1ug) using alkylation with PNBM (2.5mM).
    Lane 3 : JNK1 protein (10ng) incubated with c-jun-GST substrate (1ug) using ATP-gamma-S
    Lane 4 : JNK1 protein (10ng) incubated with c-jun-GST substrate (1ug) using ATP-gamma-S followed by alkylation with PNBM (2.5mM).


    In vitro 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 ab92570 at 1/5000, alpha-GST antibody confirms equal loading.

References

This product has been referenced in:
  • Hara M  et al. Identification of PNG kinase substrates uncovers interactions with the translational repressor TRAL in the oocyte-to-embryo transition. Elife 7:N/A (2018). Read more (PubMed: 29480805) »
  • Liu L  et al. The LIS1/NDE1 Complex Is Essential for FGF Signaling by Regulating FGF Receptor Intracellular Trafficking. Cell Rep 22:3277-3291 (2018). Read more (PubMed: 29562183) »
See all 30 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

The rabbit monoclonal antibodies (RabMAbs) can use the same anti-rabbit IgG secondary antibody that you would use with a rabbit polyclonal antibody. For Western blot, we have a line of secondary antibodies called AbExcel, which have been thoroughly tested in WB and can be used at 1:2000-1:20000 dilutions. For anti-rabbit, I'd suggest ab16284: https://www.abcam.com/donkey-rabbit-igg-hl-hrp-ab16284.html

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Answer


The main difference between ab92570 and ab133473 is the composition of the storage buffer.
The ab133473 is ideal for IP because it is supplied as cell culture supernatant with 0.01% sodium azide and the IgG concentration is at least 1 mg/ml, which is higher than ab92570.
Ab92570 can be used for IP, but it will require more antibody per reaction. This is why we launched the IP version (ab133473), which has higher concentration and no BSA in the buffer.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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