Bagby R M, Pepe F A
Histochemistry. 1978 Dec 1;58(3):219-35. doi: 10.1007/BF00495721.
Highly purified chicken gizzard myosin was used to induce antibody production in rabbits. The IgG fraction was separated from the antisera and coupled to fluorescein isothiocyanate (FITC). Specific antibody (AGM) was isolated from the IgG fraction by affinity purification. Comparisons of the specificity of IgG and AGM for chicken smooth muscle myosin revealed a much greater specificity by AGM. Staining with IgG led to an apparent cross-reactivity with guinea pig smooth muscles which was not seen with AGM staining. Therefore, staining of cells for localization of myosin was performed with AGM. Isolated cells were obtained from chicken gizzards either by collagenase digestion or by agitation of glycerinated pieces. Stained cells and cell fragments revealed the presence of myofibrils as structural units with diameters of about 1.0 micrometer. Stained myofibrils occasionally displayed regular banding patterns with a repeating period of about 1.5 +/- 0.2 micrometer. The presence of banded myofibrils in non-cultured cells shows that the organization of the contractile material is similar to that previously reported for cultured cells by Gröschel-Stewart.
使用高度纯化的鸡砂囊肌球蛋白在兔体内诱导抗体产生。从抗血清中分离出IgG组分,并与异硫氰酸荧光素(FITC)偶联。通过亲和纯化从IgG组分中分离出特异性抗体(AGM)。比较IgG和AGM对鸡平滑肌肌球蛋白的特异性发现,AGM具有更高的特异性。用IgG染色导致与豚鼠平滑肌出现明显的交叉反应,而AGM染色未出现这种情况。因此,使用AGM对细胞进行肌球蛋白定位染色。通过胶原酶消化或甘油处理切片的搅动从鸡砂囊中获得分离的细胞。染色的细胞和细胞碎片显示存在直径约为1.0微米的肌原纤维作为结构单元。染色的肌原纤维偶尔显示出规则的条纹图案,重复周期约为1.5±0.2微米。未培养细胞中带状肌原纤维的存在表明收缩物质的组织与Gröschel-Stewart先前报道的培养细胞相似。