Vallet J L
Roman L. Hruska U.S. Meat Animal Research Center, USDA, Clay Center, NE 68933.
J Anim Sci. 1994 Sep;72(9):2449-54. doi: 10.2527/1994.7292449x.
This report describes a RIA for porcine retinol binding protein (RBP) that has been validated for measurement of RBP concentrations as low as 10 ng/mL in serum, follicular fluid, uterine flushings, allantoic fluid, and endometrial and uterine tissue culture medium. The increasing displacement of porcine [125I]RBP resulting from decreasing dilutions of each sample was parallel to the standard curve, and exogenous RBP added to samples was accurately measured with the assay. To examine specificity, 1-mL samples of serum, uterine flushings, follicular fluid, and allantoic fluid were subjected to G-100 Sephadex chromatography, and the fractions were assayed for RBP with the RIA and for fluorescence, which detects retinol bound to RBP. The RBP measured with the RIA and the fluorescent fractions coeluted. Samples of uterine flushings in which RBP was measured using immunoblotting and densitometry were also assayed with the RIA. The measurements using the two methods were correlated (r = .84, P < .01). These results are evidence that the assay is specific for RBP. The limit of detection of the assay was .46 ng. The RIA should facilitate the study of RBP function and the control of its secretion by various tissues in pigs.
本报告描述了一种用于猪视黄醇结合蛋白（RBP）的放射免疫分析方法（RIA），该方法已被验证可用于测量血清、卵泡液、子宫冲洗液、尿囊液以及子宫内膜和子宫组织培养基中低至10 ng/mL的RBP浓度。每个样品稀释度降低导致猪[125I]RBP的置换增加，其与标准曲线平行，并且该分析方法能够准确测量添加到样品中的外源性RBP。为检测特异性，对1 mL血清、子宫冲洗液、卵泡液和尿囊液样品进行G-100葡聚糖凝胶柱色谱分析，并用RIA法检测各组分中的RBP，用检测与RBP结合的视黄醇的荧光法进行检测。用RIA法测定的RBP与荧光组分共洗脱。还用RIA法检测了用免疫印迹法和光密度法测定RBP的子宫冲洗液样品。两种方法的测量结果具有相关性（r = 0.84，P < 0.01）。这些结果证明该分析方法对RBP具有特异性。该分析方法的检测限为0.46 ng。该放射免疫分析方法应有助于对猪RBP功能及其在各种组织中的分泌调控进行研究。
