Affinity absorbents

ADP-Ribosyl transferase (from human placenta) [9026-30-6]. Purified by making an affinity absorbent for ADP-ribosyltransferase by coupling 3-aminobenzamide to Sepharose 4B. [Burtscher et al. Anal Biochem 152 285 1986.]... 

Lectin affinity chromatography may be used to purify glycoproteins Immobilized antibodies may be used as affinity absorbants for the antigen that stimulated their production (e.g. purification of factor VIII using immobilized anti-factor VIII antibodies)... 

ADP-Ribosyl transferase (from human placenta). Purified by making an affinity absorbent for ADP-... 

The final step in production of an affinity absorbent is removal of unbound ligand. Although the activated groups of agarose would not be expected to survive more than 12 hours it is advisable to wash the absorbent at room temperature with a O.lAf solution of pH 9.0 glycine buffer. This ensures that any possible surviving activated groups are... 

Figure 7-6. The approach of a macromolecule to an affinity absorbent for which the ligand is bound directly to the matrix (right) or bound indirectly to the matrix through an arm (left). Note that in the right figure no interaction can occur between the macromolecule and the bound ligand.

Figure 7-12. Isolation of poly A containing RNA using oligo (dT)-cellulose as the affinity absorbent. Crude rabbit reticulocyte polysomal RNA was applied to the column in a solution containing 0.01M Tris-HCl (pH 7.5) and 0.5M KCl. Nonabsorbed RNA (peak A) was eluted from the column by continued washing with the application buffer. The column was washed with a buffer of intermediate ionic strength (peak B) and finally with a buffer of low ionic strength (peak C) which removed poly A containing RNA from the column. [From H. Aviv and P. Leder, Proc. Natl. Acad. Sci. US, 69 1408 (1972).]...

Hopefully, different variations of the methodology presented here may lead to biologically important oligosaccharides, help establish enzyme speciHcities, and give access to affinity absorbents for use in isolation of saccharide-binding proteins, such as antibodies, myeloma proteins, lectins, and enzymes. 

Isol. from leaves of Hydrangea macro-phylla. Carbohydrate ligand for preparing affinity absorbents. Comly. available. 

Aminophenyl 1-thio-p-L-fucopyranoside [51885-73-5] Comly. available carbohydrate ligand for prepn. of affinity absorbents. 

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