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A2767 Sigma-Aldrich

Adenosine 5′-triphosphate–Agarose

lyophilized powder

  •  MDL number MFCD00801332

  •  eCl@ss 32160414

  •  NACRES NA.56

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Properties

Related Categories Affinity Chromatography, Affinity Resins, Core Bioreagents, Molecular Biology, Nucleotide/Coenzyme Resins,
Quality Level   200
form   lyophilized powder
extent of labeling   1-5 μmol per mL
matrix   cross-linked 4% beaded agarose
matrix activation   cyanogen bromide
matrix attachment   C-8
matrix spacer   9 atoms
storage temp.   −20°C

Description

Application

Adenosine 5′-triphosphate Agarose (5′-ATP agarose) has been used in affinity chromatography to purify uridine kinase from Ehrlich ascites tumor cells.

Packaging

1, 5, 25 mL in poly bottle

Physical form

Lyophilized powder stabilized with lactose

Safety & Documentation

Safety Information

Personal Protective Equipment 
RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number

Certificate of Origin (COO)

Please Enter a Lot Number

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
How can I hydrate Product A2767, Adenosine 5´-triphosphate-Agarose?
The resin can be hydrated by placing it in excess water, approximately 50 mL per g of resin, for at least 30 minutes.Remove the lactose stabilizer by washing the resin on a Buchner funnel with gentle vacuum, using approximately 100 mL of water per g of resin.Do not allow the resin to dry.Resuspend the resin in excess water or starting buffer to pack the column bed.
How can I elute my proteins from Product A2767, Adenosine 5´-triphosphate-Agarose?
Specifically bound proteins can be eluted with 10-100 mM ATP or ADP. Nonspecifically bound proteins can be eluted with 2 M NaCl or KCl in water or 7 M urea.
How can I regenerate Product A2767, Adenosine 5´-triphosphate-Agarose?
Since N6-[N-(6-aminohexyl)-carbamyl]-ADP is a substrate for acetate kinase and pyruvate kinase, these enzymes can be used to regenerate ATP from ADP on the resin under conditions favoring the reverse reaction. Wash the resin with 20 volumes of 50 mM Tris HCl buffer (pH 8.2), with 0.2 mM EDTA and 100 mM KCl.Incubate the resin overnight at 2-8°C in an ATP-regenerating mixture containing 0.2 mM phosphoenol pyruvate, pyruvate kinase (10 units per mL of resin), 5 mM MgCl2, 0.2 mM EDTA and 100 mM KCl in 50 mM Tris HCl buffer (pH 8.2). Wash the resin with 25 column volumes of 2 mM ATP in 2 M KCl and reequilibrate the resin with 25 column volumes of sample buffer. Alternatively, the ATP regenerating mixture can contain 20 mM acetyl phosphate, acetate kinase (6.8 units per mL of resin), and 3 mM MgCl2 in 100 mM Tris HCl (pH 7.6). If acetate kinase is used, prewash the resin with 25 column volumes of 100 mM Tris HCl (pH 7.6), but use the same final wash and reequilibration steps as given for pyruvate kinase.
Can I store Product A2767, Adenosine 5´-triphosphate-Agarose, after I have hydrated it?
For the short term, it is possible to store the hydrated resin in 20% ethanol and dilute buffer at 2-8°C.In general, the hydrated resin can be stored refrigerated in water or buffer containing a bacteriostat, such as 0.02% sodium azide or thimerosal. Do not autoclave or freeze the hydrated resin. The resin can be used several times without loss of effectiveness. However, the ATP will slowly hydrolyze to ADP over time, and under certain conditions this process may be accelerated during usage.The resin can be regenerated; please see the regeneration FAQ for this product.
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Protocols & Articles
Peer-Reviewed Papers
15

References

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