小鼠VEGF-C ELISA试剂盒#产品技术说明书

试剂盒采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。往预先包被小鼠血管内皮生长因子C（VEGF-C）捕获抗体的包被微孔中，依次加入标本、标准品、HRP标记的检测抗体，经过温育并洗涤。用底物TMB显色，TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成终的黄色。颜色的深浅和样品中的小鼠血管内皮生长因子C（VEGF-C）呈正相关。用酶标仪在450nm 波长下测定吸光度（OD 值），计算样品浓度。

标准品浓度依次为：320、160、80、40、20、10 pg/mL

试剂盒性能

1. 检测范围：10 pg/mL – 320 pg/mL。

2. 灵敏度：检测浓度小于1.0 pg/mL。

3. 特异性：不与其它可溶性结构类似物交叉反应。

4. 重复性：板内变异系数小于10% ，板间变异系数小于15% 。

ASSAY PROCEDURE

1. Prepare all reagents before starting assay procedure. It is recommended that all Standards ａnd Samples be added in duplicate to the Microtiter plate.

2. Add 50μl of Standard or Sample to the appropriate wells. Blank well doesn’t add anyting.

3. Add 100μl of Enzymeconjugate to standard wells and sample wells except the blank well, cover with an adhesive strip and incubate for 60 minutes at 37°C.

4. Wash the Microtiter Plate 4 times.

Manual Washing - Remove incubation mixture by aspirating contents of the plate into a sink or proper waste container. Using a squirt bottle, fill each well completely with Wash Solution (1X), then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure for a total of four times. After final wash, invert plate, ａnd blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame.

Automated Washing - Aspirate all wells, then wash plates four times using Wash Buffer (1X). Always adjust your washer to aspirate as much liquid as possible ａnd set fill volume at 350μL/well/wash. After final wash, invert plate, ａnd blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears.

5. Add Substrate A 50μl ａnd Substrate B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.

6. Add 50μl S Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.

7. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.

原创作者：上海杰普斯生物科技有限公司