Gearhart J, Oster-Granite M L, Musser J M
Biochem Genet. 1981 Jun;19(5-6):445-55. doi: 10.1007/BF00484617.
There are two common electrophoretic variants (allozymes) of murine glucosephosphate isomerase (GPI). In order to develop immunocytochemical procedures that are allozyme-specific, the two variants (GPI-1A and GPI-1B) were purified from skeletal muscle of several mouse strains and used as antigens for the elicitation of antibodies. The allozymes were purified to a specific activity of 800 units/mg by substrate elution from cellulose phosphate. When the purified allozymes were presented as antigens to goats, rabbits, and mice (both syngeneically and allogeneically), the goats and rabbits produced high titers of anti-GPI antibody, but no humoral antibody was detected in the mice, as determined by radioimmunoassays. Antisera specific for the GPI-1B variant were enriched by absorbing selected sera with GPI-1A conjugated to Sepharose 4B. No antisera specific for GPI-1A were detected following the immunizations. The specificity of the anti-GPI-1B antisera provides a unique opportunity for the development of immunocytochemical procedures for studying the distribution fo this allozyme in chimeric mouse tissues.
小鼠葡萄糖磷酸异构酶（GPI）有两种常见的电泳变体（别构酶）。为了开发具有别构酶特异性的免疫细胞化学方法，从几种小鼠品系的骨骼肌中纯化出这两种变体（GPI-1A和GPI-1B），并将其用作抗原以引发抗体。通过从磷酸纤维素上进行底物洗脱，将别构酶纯化至比活性为800单位/毫克。当将纯化的别构酶作为抗原分别给予山羊、兔子和小鼠（同基因和异基因）时，山羊和兔子产生了高滴度的抗GPI抗体，但通过放射免疫测定法测定，在小鼠中未检测到体液抗体。通过用与琼脂糖4B偶联的GPI-1A吸收选定的血清，富集了对GPI-1B变体特异的抗血清。免疫后未检测到对GPI-1A特异的抗血清。抗GPI-1B抗血清的特异性为开发用于研究这种别构酶在嵌合小鼠组织中分布的免疫细胞化学方法提供了独特的机会。
