Antibodies proteolytic digestion

Mazzoni, M. R., Malinski, J. A, and Hamm, H E. (1991) Structural analysis of rod GTP-binding protein, Gt-limited proteolytic digestion pattern of Gt with four proteases defines monoclonal antibodies epitope J Biol. Chem 266,14,072—14,081... 

The process of antibody optimization also requires that different temperatures of antigen retrieval be explored, and although citrate buffer at 10 mMol/litre is a good universal retrieval solution, a higher pH may be required for more capricious antigens as previously discussed (Shi et al, 1995). In addition, the synergistic action of proteolytic digestion should be routinely explored as a pre-MW and post-MW procedure. 

No direct three-dimensional structure determination of any GPCR bus been carried out yet However, die structure of the membrane-embedded helices of bucteriorhodopsin has been solved by electron crystallography [131]. Experimental data from deletion mutations, antibody targeting, and proteolytic digestion experiments provide evidence (hat the overall features of GPCRs and... 

Singh and Preiss265 concluded that, although some homology exists between the three starch-branching enzymes, there are major differences in the structure of BEI when compared to BEIIa and BEIIb, as shown by its different reactivity with some monoclonal antibodies and differences in amino acid composition and proteolytic digest maps. 

Epitope excision was performed by application of 2-5 xg antigen to the antibody microcolumn (Tian et al. 2005). Binding was performed for 60 min at 20°C. After washing with binding buffer, proteolytic digestion was carried out on the column with 0.2 xg protease in 200 xl PBS for 2 h at 37°C. Supernatant unbound peptides were removed using washing buffer and the epitope dissociated by addition of 500 pi 0.1 % trifluoroacetic acid (TFA). After incubation for 15 min at 20 C, the epitope eluate was lyophilised and reconstituted in 10 pi NaAc buffer. 

Mechanism of Action. TSH and LATS cause similar effects on glucose oxidation, P uptake (F2), and iodine release (E6). The time course is delayed, presumably due to the molecular size of the LATS. Antihuman TSH antibody did not inhibit the effect of LATS on P uptake into phospholipids or on glucose oxidation. As stated above (M13) proteolytic digestion of LATS makes it a short-acting thyroid stimulator. 

Although sensitivity may be increased in many instances using polymer-based labels, others have reported that with some antibodies, particularly those that require proteolytic digestion or target unmasking, the EnVision method revealed slightly weaker staining and less sensitivity. This outcome was believed... 

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