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Pierce Monomeric Avidin Resins and Kit


Ideal affinity support for gentle, reversible binding of biotinylated proteins.

Pierce Monomeric Avidin Kit and ResinsThe Thermo Scientific Pierce Monomeric Avidin Kit and Resins enable gentle, non-denaturing affinity purification of biotinylated molecules.

The Immobilized Monomeric Avidin protein binds biotin with high specificity and moderate affinity, allowing non-biotin molecules to be washed away and then the bound biotin-labeled molecules to be competitively eluted using 2mM biotin in phosphate buffered saline (PBS). This technique provides the gentlest elution conditions for biotinylated protein purification and avoids the contamination and other problems associated with traditional avidin and streptavidin methods. The monomeric avidin column can then be regenerated by using 0.1M glycine to strip the column of residual bound biotin without losing the ability to bind another biotinylated sample.

Highlights:

  • Non-denaturing – purifies biotinylated products using mild elution conditions (2mM free biotin)
  • Reusable – Monomeric Avidin Agarose can be regenerated and reused at least 10 times, with only a marginal loss in biotin binding capacity (approximately 2.5% decrease per regeneration)
  • Specific – retains biotin-binding specificity and low nonspecific binding of native avidin
  • Flexible – bind samples in a variety of physiological buffers and elute either with 0.1M glycine or by competition with 2mM biotin
  • Good binding capacity – greater than 1.2mg biotinylated BSA/mL resin
  • Convenient – purchase standalone agarose or UltraLink Resins or a complete purification kit with prepacked column and buffers

Product Details:

With typical avidin or streptavidin, the biotin-binding affinity (Kd = 10-15M) is so great that purification with these traditional media require denaturing conditions for elution, such as 8M Guanidine•HCl at pH 1.5 or boiling in reducing SDS-PAGE sample loading buffer. In addition to their adverse effects on the biotinylated protein of interest, such harsh elution conditions typically cause extraneous proteins (i.e., the avidin or streptavidin subunits) to strip from the resin and co-elute with the desired purification product. By contrast, when avidin is coupled to a resin (e.g., crosslinked beaded agarose or UltraLink Resin) as the subunit monomer, its specificity for biotin is retained, but its biotin-binding affinity decreases to a level (Kd = 10-8M) that is conducive to subsequent elution.

Biotin binding and elution with Monomeric Avidin
Biotin binding and elution with Monomeric Avidin Agarose.

References:

  1. Petrotchenko E. V. et al. (2011) An isotopically coded cid-cleavable biotinylated cross-linker for structural proteomics. Mol. Cell. Proteomics. 10, M110.001420-.
  2. Friis S. et al. (2011) Transport via the transcytotic pathway makes prostasin available as a substrate for matriptase. J. Biol. Chem. 286, 5793-802.
  3. Nie Y. and Kaback H. R. (2010) Sugar binding induces the same global conformational change in purified lacy as in the native bacterial membrane. PNAS. 107, 9903-8.
  4. Forejtnikova H. et al. (2010) Transferrin receptor 2 is a component of the erythropoietin receptor complex and is required for efficient erythropoiesis. Blood. 116, 5357-67.
  5. Feria Bourrellier A. B. et al. (2010) Chloroplast acetyl-coa carboxylase activity is 2-oxoglutarate-regulated by interaction of pii with the biotin carboxyl carrier subunit. PNAS. 107, 502-7.

Related Products:

Biotin
Biotin Labeling Reagents and Kits
Cell Surface Protein Isolation Kit


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
20227 Pierce Monomeric Avidin Agarose Kit, 2mL
Formulation: Multi-component kit
Sufficient For: Binding ≥ 2.4mg biotinylated BSA per column

Kit Contents:
Pierce Monomeric Avidin Column, 2mL, 1 column
PBS Pack (makes 500mL), 1 pack
Biotin Binding and Elution Buffers, 200mL
Regeneration Buffer, 250mL
Column Accessories
1-column kit Product Instructions for product #20227 Pierce Monomeric Avidin Agarose Kit, 2mL MSDS for product #20227 Pierce Monomeric Avidin Agarose Kit, 2mL Certificate of Analysis for product #20227 Pierce Monomeric Avidin Agarose Kit, 2mL $356.00

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20228 Pierce Monomeric Avidin Agarose
Formulation: Avidin monomers on crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Binding ≥ 1.2mg biotinylated BSA per mL of resin
5mL Product Instructions for product #20228 Pierce Monomeric Avidin Agarose MSDS for product #20228 Pierce Monomeric Avidin Agarose Certificate of Analysis for product #20228 Pierce Monomeric Avidin Agarose $384.00

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20267 Pierce Monomeric Avidin Agarose
Formulation: Avidin monomers on crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Binding ≥ 1.2mg biotinylated BSA per mL of resin
10mL Product Instructions for product #20267 Pierce Monomeric Avidin Agarose MSDS for product #20267 Pierce Monomeric Avidin Agarose Certificate of Analysis for product #20267 Pierce Monomeric Avidin Agarose $625.00

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20228B Pierce Monomeric Avidin Agarose
Formulation: Avidin monomers on crosslinked 4% beaded agarose, slurried in water with sodium azide
Sufficient For: Binding = 1.2mg biotinylated BSA per mL of resin
Custom MSDS for product #20228B Pierce Monomeric Avidin Agarose Certificate of Analysis for product #20228B Pierce Monomeric Avidin Agarose
53146 Pierce Monomeric Avidin UltraLink Resin
Formulation: Avidin monomers on UltraLink Resin, slurried in water with sodium azide
Sufficient For: Binding ≥ 1.2mg biotinylated BSA per mL of resin
5mL Product Instructions for product #53146 Pierce Monomeric Avidin UltraLink Resin MSDS for product #53146 Pierce Monomeric Avidin UltraLink Resin Certificate of Analysis for product #53146 Pierce Monomeric Avidin UltraLink Resin $433.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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