LIT195B 6/17/98 11:37 AM Page Cvr2 Bio-Beads® SM Hydrophobic and Polar Interaction Adsorbents Instruction Manual Bio-Rad Laboratories, 2000 Alfred Nobel Dr.
LIT195B 6/17/98 11:37 AM Page i Table of Contents Section 1 Introduction ..........................................1 Section 2 Product Description..............................2 Section 3 Instructions for Use ..............................4 3.1 Preparation of Bio-Beads SM Adsorbent (Bulk) ............................................4 Column Method ..............................................4 3.2 Section 4 Sample Protocols...................................7 4.1 4.2 4.3 4.
LIT195B 6/17/98 11:37 AM Page 1 Section 1 Introduction Bio-Beads SM adsorbents, for adsorbing organics from aqueous solutions, are neutral, macroporous polymeric Analytical Grade adsorbents of high surface area, which are intended for laboratory use only. The adsorbent is composed of a large number of highly crosslinked microspheres. This macroreticular structure gives it high surface area and uniform pore sizes.
LIT195B 6/17/98 11:37 AM Page 2 Table 1. Description of Bio-Beads SM Adsorbents Bio-Beads SM-2 Adsorbent Chemical nature Polystyrene-divinyl-benzene Polarity Nonpolar Dipole moment 0.3 Debye detergents such as deoxycholate,4-6 NP-40,7 and emulgen 911.8-10 Bio-Beads adsorbents have been used to separate water soluble steroids, 11,12 phenols, 13 drugs, 14,15 pesticides,16-20 trace organics,21-23 and rhodamine.24 Table 2 lists some other general applications of BioBeads adsorbents.
LIT195B 6/17/98 11:37 AM Page 4 A complete list of more than 100 applications for Bio-Beads adsorbents can be obtained by requesting technical bulletin 1461. 2. Section 3 Instructions for Use 3.1 Preparation of Bio-Beads SM Adsorbent (Bulk) Due to the macroporous nature of the adsorbent, air will sometimes become trapped within the pores. As a result, it will float in solution. To correct this, slurry the adsorbent in the solution you will use later.
LIT195B 6/17/98 11:37 AM Page 6 3.3 Batch Method The batch method is the addition of adsorbent directly into the sample followed by stirring to achieve the adsorption. This is a very simple and common method, though it requires 90–120 minutes, as opposed to 15 minutes for the column procedure. 1. Weigh 5 g of Bio-Beads SM adsorbent for every 25 ml of solution. 2. Add the adsorbent to the solution. 3. Use either a stir bar or a mechanical agitator to mix the solution at room temperature for 2 hours. 4.
LIT195B 6/17/98 11:37 AM Page 8 3. Wash with 10 mM phosphate buffer at 0.03 ml/min. Collect fractions and assay for protein. The column can be regenerated by washing in 4 bed volumes of methanol, followed by rinsing in DI water. 4.2 Adsorption of Unconjugated Fluorescent Dye This procedure was used by Spack et al. to reduce high background fluorescence with rhodamine-labeled antibodies.24 It should be useful for many fluorescent labels.
LIT195B 6/17/98 11:37 AM Page 10 Section 5 Storage All Bio-Beads SM adsorbents are stable for 5 years when stored at room temperature in the unopened container. Section 6 References 8. Gibson, G. G. and Schenkman, J. B., J. Biol. Chem., 253, 5957 (1978). 9. Warner, M., J. Biol. Chem., 257 (21), 12995 (1982). 10. Bruch, R. C., Thotakura, N. R. and Bahl, O. P., J. Biol. Chem., 261 (20), 9450 (1986). 11. Shackleton, C. H. L., Sjovall, J. and Wisen, O., Clin. Chim. Acta, 27, 354 (1970). 12. Bradlow, H.
LIT195B 6/17/98 11:37 AM Page 12 20. Young, C., Proc. Nat. Sci. Counc. ROC (B), 8 (2), 119 (1984). 21. Wigilius, B., Boren, H., Carlberg, G. E., Grimvall, A., Lundgren, B. V. and Savenhed, R., J. Chromatog., 391, 169 (1987). 22. Deshmukh, S. W. and Pangarkar, V. G., Indian Chem. Eng., 26 (3), 35 (1984). 23. Libbey, A. J. Jr., Analyst, 3, 1221 (1986).
