New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates

Purification of the sodium transport enzyme oxaloacetate decarboxylase by affinity chromatography on avidin sepharose

“…This enzyme also Membranes were prepared from Klebsiella aerogenes grown anaerobically on citrate as in [3,4]. Oxaloacetate decarboxylase was solubilized from these membranes with 1.5% Triton X-100 in 2 mM K-phosphate buffer (pH 7.0) and applied to a column of monomeric avidin-Sepharose [9]. The column was washed with -6 vol.…”

“…Protein was determined as in Because of the very high affinity of avidin for biotin (K = IO-l5 M) [ 131 it has not been possible to recover biotincontaining enzymes from avidin affinity columns under non-denaturing conditions. Based on an observation that the monomeric form of avidin has a reduced affinity toward biotin [ 141, several groups have developed monomeric avidin affinity columns for the purification of biotincontaining proteins [9,15,16].…”

Metamorphosis‐dependent transcriptional regulation of xak‐c, a novel Xenopus type I keratin gene

“…When these peptides were chemically synthesized, cysteine residues were additionally bound to the N-terminus of each of the two peptides, because the cysteine residue is necessary to bind the peptides to keyhole limpet hemocyanin (Liu et al, 1979). The antibodies against these peptides were generated according to Suzuki et al (2002).…”

Idiotypic and subgroup analysis of human monoclonal rheumatoid factors. Implications for structural and genetic basis of autoantibodies in humans.

“…The chosen prototype subgroup sequences were distinct; the greatest homology was only 50% between PHII and PHIII. The peptides were coupled to keyhole limpet hemocyanin (KLH) through a terminal cysteine moiety, using m-maleimido-benzoyl-N-hydroxysuccinamide ester (21 Immunoblot analysis. The binding specificity of antipeptide antisera and antibody 17.109 were also assessed by an immunoblot method, as previously described (14,18,22).…”