g-linked ATP agarose resin

g-linked ATP agarose resin Summary

• Description: Affinity resins have been widely used for the purification of enzymes that bind nucleotides and related molecules. Resins in which ATP is linked via the gamma-phosphate have been valuable in identifying proteins in the purine-binding proteome, which includes kinases, heat shock proteins and other ATP-binding proteins.
  
  ATP Agarose resin comprises ATP attached to agarose beads via its gamma-phosphate. Two forms of the resin are available with low and high ligand substitution. A long hydrophilic spacer (14-atom) is used to minimise unwanted hydrophobic interactions and to facilitate unhindered interactions with biomolecules.
  
  The Control Resin is just the agarose with no ATP bound.
• Specificity: Affinity resins have been widely used for the purification of enzymes that bind nucleotides and related molecules. Resins in which ATP is linked via the gamma-phosphate have been valuable in identifying proteins in the purine-binding proteome, which includes kinases, heat shock proteins and other ATP-binding proteins.
  
  ATP Agarose resin comprises ATP attached to agarose beads via its gamma-phosphate. Two forms of the resin are available with low and high ligand substitution. A long hydrophilic spacer (14-atom) is used to minimise unwanted hydrophobic interactions and to facilitate unhindered interactions with biomolecules.
  
  The Control Resin is just the agarose with no ATP bound.

Applications/Dilutions

• Application Notes: High (8-12 umol/ml) - 2ml

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.

Notes

This product is manufactured by Abcam and distributed by Novus Biologicals.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. This product is guaranteed for 1 year from date of receipt and this statement overrides any mentioned guarantee period on the limitations section of this products datasheet. Please contact technical@novusbio.com with questions.

Background

ATP linked to a support via its g-phosphate has been used for affinity purification of kinases, heat-shock proteins and other ATP-binding proteins. KinaseBind is a g-phosphate-linked ATP resin with a long hydrophillic spacer that facilitates molecular interactions with biomolecules. Linkage of ATP via the terminal phosphate means that the immobilised ligand cannot be removed by phosphatases in crude extracts.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are guaranteed for 6 months from date of receipt.

Publications for g-linked ATP agarose resin (510-0002)(8)

Publications using 510-0002

• Backe SJ, Sager RA, Heritz JA et al. Activation of autophagy depends on Atg1/Ulk1-mediated phosphorylation and inhibition of the Hsp90 chaperone machinery Cell reports 2023-07-25 [PMID: 37453059]
• Baker-Williams AJ, Hashmi F, Budzy?ski MA et al. Co-chaperones TIMP2 and AHA1 Competitively Regulate Extracellular HSP90:Client MMP2 Activity and Matrix Proteolysis Cell Reports 2019-08-13 [PMID: 31412254] (, )
• Sager RA, Woodford MR, Backe SJ et al. Post-translational Regulation of FNIP1 Creates a Rheostat for the Molecular Chaperone Hsp90 Cell Rep 2019-01-29 [PMID: 30699359]
• Mark R Woodford, Rebecca A Sager, Elijah Marris, Diana M Dunn, Adam R Blanden, Ryan L Murphy, Nicholas Rensing, Oleg Shapiro, Barry Panaretou, Chrisostomos Prodromou, Stewart N Loh, David H Gutmann, Dimitra Bourboulia, Gennady Bratslavsky, Michael Wong, Mehdi Mollapour Tumor suppressor Tsc1 is a new Hsp90 co‐chaperone that facilitates folding of kinase and non‐kinase clients The EMBO Journal 2017-11-10 [PMID: 29127155]
• Rao R, Fiskus W, Yang Y et al. HDAC6 inhibition enhances 17-AAG--mediated abrogation of hsp90 chaperone function in human leukemia cells. Blood. 2008-09-01 [PMID: 18591380]
• Duncan JA, Bergstralh DT, Wang Y, Willingham SB, Ye Z, Zimmermann AG, Ting JP. ryopyrin/NALP3 binds ATP/dATP, is an ATPase, and requires ATP binding to mediate inflammatory signaling. Proc Natl Acad Sci U S A. 104(19):8041-6. 2007-05-08 [PMID: 17483456]
• Harthill JE, Meek SE, Morrice N, Peggie MW, Borch J, Wong BH, Mackintosh C. Phosphorylation and 14-3-3 binding of Arabidopsis trehalose-phosphate synthase 5 in response to 2-deoxyglucose. The Plant Journal 47 (2), 211-223. 2006-07-01 [PMID: 16771775]
• Jaleel M, Saha S, Shenoy AR, Visweswariah SS. The kinase homology domain of receptor guanylyl cyclase C: ATP binding and identification of an adenine nucleotide sensitive site. Biochemistry;45(6):1888-98. 2006-02-14 [PMID: 16460035]

FAQs for g-linked ATP agarose resin (510-0002). (Showing 1 - 1 of 1 FAQs).

1. What is the binding capacity of the ATP Agarose?
   • The binding of any ATP-binding protein to our resin will depend on a number of factors. These include amongst others: the formulation of the binding buffer, the incubation time of the sample with the resin (flow rate), the concentration of the ATP-binding protein, the affinity of the binding protein for ATP and the size of the binding protein (steric hindrance can be a factor with larger proteins). The only true way to establish the capacity of the column for any individual binding protein is by experimentation. If you require mg scale purification then our ATP agarose should be suitable, especially as the resin can be reused. The unique nature of our ATP resin is that it is not subject to dephosphorylation like many other similar resins available from other suppliers. This means that you can simply wash, re-equilibrate and reuse the resin multiple times. This will enable you to obtain mg quantities of your protein even if it takes more than one purification.