Receptor purification

Two steroid affinity resins have proven to be most useful in the purification of PR. One, developed by Grandics et al. [17], is a deoxycorticosterone derivative modified through the 21-carbon and linked to Sepharose 2B through an epoxide and a 

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Estrogen receptors, purification, 3 845 Estrogens, synthetic, 13 3 Estuarine environment, 26 20-21 p-carbides, 4 691, 692... 

Due to the strong and selective binding that characterizes many affinity ligands, solutes that are analyzed or purified with these ligands can often be separated with little or no interferences from other sample components. In many cases, the solute of interest can be isolated in only one or two steps, with purification yields of hundred- to several thousandfold being common [2,4-6]. In work with hormone receptors, purification yields approaching one millionfold have even been reported with affinity-based separations [5]. 

Novick, D., P. Orchansky, M. Revel, and M. Rubinstein, The human interferon-gamma receptor. Purification, characterization, and preparation of antibodies. J Biol Chem, 1987. 262(18) 8483-7. 

Regnier FE (1984) HPLC of membrane proteins. In Receptor Purification Procedures, Vol. 2, JC Venter and LC Harrison, Eds. Liss, New York. 

Stein M, Meyer HE, Hasilik A, von Eigura K. 46-kDa mannose 6-phosphate-specific receptor purification, subunit composition, chemical modification. Biol. Chem. Hoppe-Seyler 1987 368 927-936. 

Neurotransmitter and Hormone Receptors Purification by Affinity Terry M. Phillips... 

Novick, D. (1987). The Human Interferon-y-receptor Purification, Characterization, and Preparation of Antibodies, Biol. Chem. 262 8483-8487. 

It could happen that they were working on a receptor purification, only to read after one year of intensive efforts that the cDNA had been expression cloned. With cDNA, you could simply do a lot more than with purified protein. You got the entire sequence. You could change the sequence, express the protein, produce antibodies against the entire protein or parts of it, measure the function, and so on. 

Grisshammer R. and Tucker J. 1997. Quantitative evaluation of neurotensin receptor purification by immobilized metal affinity chromatography. Protein Expr. Purif., 11, 53. 

Lomasney, J.W., Leeb-Lundberg, L.M., Cotecchia, S., Regan, J.W., DeBemardis, J.F., Caron, M.G. and Lefkowitz, R.J. (1986). Mammalian aj-adrenergic receptor. Purification and characterization of the native receptor ligand binding subunit. /. Biol Chem., 261, 7710-7716... 

Reliable and reproducible procedures are needed to purify auxin receptor protein for antibody production and for further biochemical characterization. Two auxin affinity methods have been reported [11,17], but such methods have yielded indifferent results in our hands [22]. This communication describes auxin receptor purification using commercially available chromatographic materials and the initial characterization of monoclonal and polyclonal antibodies produced against the resulting preparations. 

Receptor Purification Ail except the elution step are performed at 4°C. 

The recovery was 50-75%, with respect to the amount of receptor present in the aggregate-free preparation (depending on the initial quality of the calf uterus tissue), and the purity was estimated to about 80%. These results were quite competitive with those afforded by traditional affinity chromatography [22,23], if not better, and were obtained in a short time without previous experience in the field of estrogen receptor purification. The quantity of material that could be treated by this technique was only limited by the size of the gel filtration columns. On the other hand, the water-soluble macroligand could be recovered in the void volume of the column, at the second filtration step, and recycled for further purification experiments, after removal of the non-specifically bound contaminants. 

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