Direct Antibody Method for Formalin Fixed, Paraffin Embedded Renal Biopsies: A Comparison With the Peroxidase Labeled Streptavidin/Biotin Method
Direct Antibody Method for Formalin Fixed, Paraffin Embedded Renal Biopsies: A Comparison With the Peroxidase Labeled Streptavidin/Biotin Method
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用于福尔马林固定、石蜡包埋肾活检的直接抗体方法：与过氧化物酶标记的链霉亲和素/生物素方法的比较
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发表时间：
1996
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通讯作者：
I. Birchall
中科院分区：
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作者：
I. Birchall
本文比较了过氧化物酶标记抗体直接法和过氧化物酶标记链霉亲和素/生物素（LSAB）法对中性缓冲、福尔马林固定、石蜡包埋肾活检组织中免疫球蛋白A、G、M、补体（C3 c）、纤维蛋白原、κ和λ轻链的检测。这些被脱蜡，再水化，封闭内源性过氧化物酶，和0.025%的蛋白酶VIII型消化3分钟。一抗的检测效率进行了测定，通过棋盘滴定，和最佳稀释度的每种抗体在室温下应用1小时。两种方法均用3，3 ′-二氨基联苯胺四盐酸盐（DAB）检测抗体结合位点，两种方法检测到的免疫反应物在每个活检组织中的分布和数量相似。结果表明，LSAB对各抗体均具有较高的检测效率，且反应产物的强度均较LSAB高。
Abstract This report compares a direct, peroxidase conjugated antibody method with a peroxidase labeled streptavidin/biotin (LSAB) method for the detection of immunoglobulins A, G and M, complement (C3c), fibrinogen and kappa and lambda light chains in neutral buffered, formalin fixed, paraffin embedded renal biopsy tissue.Twenty-five percutaneous renal biopsies were selected and 2 μm serial sections cut. These were deparaffinized, rehydrated, blocked for endogenous peroxidase, and digested with 0.025% protease type VIII for 3 min. Detection efficiencies of the primary antibodies were determined by checkerboard titrations, and optimal dilutions of each antibody were applied for 1 hr at room temperature. Antibody binding sites in both methods were detected with 3,3′diaminobenzidine tetrahydrochloride (DAB).Both methods detected similar distributions and quantities of immune reactants in each biopsy. A greater detection efficiency was achieved for each antibody with LSAB, and the intensity of the reaction p...