Capel P J, Gerlag P G, Hagemann J F, Koene R A
J Immunol Methods. 1980;36(1):77-80. doi: 10.1016/0022-1759(80)90096-4.
The effect of reduction by 2-mercaptoethanol (2-ME) on antibody activity was studied in antisera with high and low IgG concentrations. Sera obtained from B10.LP nu/nu mice during a primary response against a rat PVG/c skin graft contained only 2-ME sensitive antibodies. However, when analyzed on sucrose gradients, IgG as well as IgM antibody activity was present. After raising the low serum IgG concentration (0.3-0.7 mg/ml) of these sera by addition of normal mouse serum (5.1 mg IgG/ml), 2-ME resistant antibodies became detectable. Hyperimmune C57BL6 anti PVG/c lymphocyte serum with a high IgG concentration (20.2 mg/ml) and antibody activity predominantly located in the IgG class was not affected by 2-ME treatment. These data show that IgG antibodies, although less susceptible to reduction than IgM antibodies, are not resistant to this treatment. At high IgG concentrations the proportion of inactivated IgG with specific antibody activity will be negligible but at low IgG concentrations the use of this method leads to serious underestimation of IgG antibody activity.
研究了2-巯基乙醇（2-ME）还原对高IgG浓度和低IgG浓度抗血清中抗体活性的影响。在B10.LP裸鼠对大鼠PVG/c皮肤移植的初次免疫反应期间获得的血清中仅含有对2-ME敏感的抗体。然而，当在蔗糖梯度上进行分析时，存在IgG以及IgM抗体活性。通过添加正常小鼠血清（5.1mg IgG/ml）提高这些血清的低血清IgG浓度（0.3-0.7mg/ml）后，可检测到对2-ME有抗性的抗体。高IgG浓度（20.2mg/ml）且抗体活性主要位于IgG类别的超免疫C57BL6抗PVG/c淋巴细胞血清不受2-ME处理的影响。这些数据表明，IgG抗体虽然比IgM抗体更不易被还原，但对这种处理并非具有抗性。在高IgG浓度下，具有特异性抗体活性的失活IgG比例可忽略不计，但在低IgG浓度下，使用该方法会导致对IgG抗体活性的严重低估。
