Antibody—deglycosylated

Dissolve deglycosylated ricin A chain (dgA) in 0.1 M sodium phosphate, 0.15 M NaCl, 10 mM EDTA, pH 7.5, at a concentration of 10 mg/ml. The buffer should be degassed under vacuum and nitrogen bubbled through it to remove oxygen. Prepare half the amount of A-chain solution as the amount of antibody prepared... 

Ghetie, V., Ghetie, M.-A., Uhr, J. W., and Vitetta, E. S. (1988) Large scale preparation of immunotoxins constructed with the Fab fragment of IgGl murine monoclonal antibodies and chemically deglycosylated ricin A chain. /. Immunol. Methods 112, 267—277. 

Rebe S, Solomon B. Deglycosylation of anti-beta amyloid antibodies inhibits microglia activation in BV-2 cellular model. Am J Alzheimers Dis Other Demen 2005 20(5) 303-13. 

The involvement of the proteasome suggests that the MHC class I heavy chains migrate back to the cytosol. Indeed, a cytosolic disposition of the deglycosylated intermediate can be confirmed by subcellular fractionation experiments. Furthermore, this intermediate can be immunoprecipitated from material reactive with anti-proteasome antibodies. Apparently, US2 and USll induce retrograde translocation, or dislocation , of MHC class I heavy chains from the ER to the cytosol, where the heavy chains are subjected to proteolytic degradation by the proteasome (Wiertz et al. 1996a,b). 

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