Thermo Scientific Pierce Boronic Acid Resin enables ribonucleotide and oligonucleotide RNA isolation.
The immobilized boronic acid is an easy-to-use polyacrylamide affinity support for purification of ribonucleotides and other small molecular weight compounds that contain cis-diol groups. The ligand (m-aminophenyl-boronic acid) binds to the cis-diol groups on the sugar portion of nucleotides, forming a reversible five-member ring complex. After washing away non-bound molecules from the sample, the complex can be dissociated and the nucleotide eluted by lowering the pH or by addition of sorbitol.
- Ligand: boronic acid
- Support: beaded polyacrylamide (1800 MW exclusion limit)
- Spacer: m-aminophenyl group
- Loading: 100µmol boronate per milliliter of settled resin
- Capacity: at least 99% binding and recovery of 110µmol AMP per milliliter of settled resin
- Format: 50% resin slurry in 0.05% sodium azide
The polyacrylamide support used in this product excludes molecules greater than 1800 MW from entering the internal spaces of the beads, allowing only smaller molecules to interact and bind with the full measure of boronate ligand. Consequently, the resin may not purify glycosylated proteins efficiently, although it has been used successfully for this application. Boronic acid affinity chromatography has been used to isolate ribonucleoside, to purify nucleosidyl peptide, separate ribonucleosidases in tissue extracts, separate RNA and oligoribonucleotide, and determine of the amount of non-enzymatic glycosylation present in peripheral nerve from diabetic and control rats and dogs.
|Chemical structure of Thermo Scientific Pierce Boronic Acid Resin.
- Gehrke, C.W., et al. (1978). J. Chromatogr. 150: 455-476.
- Hase, S., et al. (1992). Biosci. Biochem. 56(10): 1676-1677.
- Mallia, A.K., et al. (1981). Anal. Letters 14(B8): 649-661.
- Vlassara, H., Brownlee, M. and Cerami, A. (1981). Proc. Natl. Acad. Sci. USA, 78: 5190-5192.
- Londono, I. and Bendayan, M. (2001). Am. J. Physiol. Renal. Physiol. 280: F103-11.
- Annamalai, A.E., et al. (1979). Anal. Biochem. 99: 85-91.
- Olsson, R.A. (1979). J. Chromatogr. 176: 239-41.
- Pace, B. and Pace, N.R. (1980). Anal. Biochem. 107: 128-35.
Review of affinity purification