人α肌动蛋白(αActin)ELISA检测试剂盒子科生物现货供应,凡购买子科生物子科生物任何一款ELISA酶联免疫分析检测试剂盒,都可以享受免费代测服务,外地客户可以由技术老师的指导下正确放置好样本的保存方式后快递邮寄到我公司技术部,本地客户可以享受免费上门取样服务!
[产品名称]人α肌动蛋白(αActin)ELISA检测试剂盒
[英文名称]Human Alpha-Actin,α Actin ELISA Kit
[货号]ZK-H1663
[试剂盒]保存条件、保质期2-8℃低温保存下,6个月的保质期。
[运输方式]当地客户可以由专门配送人员免费送货上门货快递,外地客户当天下单当天免费快递送货上门。
[技术服务]凡购买子科生物子科生物任何一款ELISA酶联免疫分析检测试剂盒,都可以享受免费代测服务,外地客户可以由技术老师的指导下正确放置好样本的保存方式后快递邮寄到我公司技术部,本地客户可以享受免费上门取样服务!
人α肌动蛋白(αActin)ELISA检测试剂盒实验测定具体步骤:
1.确定本次检测所需的已包被抗体的酶标板孔数目,并增加 1 孔作为 TMB 空白显色孔。 总数=样品数+9;做双份检测时×2。其余重包装好放如冰箱中。
2.将标准品各 0.1ml 依次加入一排 7 孔中,1 孔只加样品稀释液的作为零孔。对于血清、体液、 组织匀浆或细胞培养上清,直接加已用样品稀释液稀释的样品 100μ ι 。
3.酶标板加上盖,37℃反应 90 分钟。
4.反应后用自动洗板机吸去酶标板内的液体;或甩去酶标板内液体,再对着吸水纸拍几下。 不洗。
5.将准备好的生物素抗体工作液按每孔 0.1ml 依次加入。(TMB 空白显色孔除 外)。37℃反应 60 分钟。
6. 0.01M TBS 或 0.01M PBS 洗涤 3 次,每次浸泡 1 分钟左右。
7.将准备好的 ABC 工作液按每孔 0.1ml 依次加入(TMB 空白显色孔除外)。37℃反应 30 分钟。
8. 0.01M TBS 或 0.01M PBS 洗涤 5 次,每次浸泡 1-2 分钟左右。
9.按每孔 90μ ι 依次加入已在 37℃平衡 30 分钟的 TMB 显色液,37℃避光反应 20-25 分钟 (注意:显色时间供参考,因用户实验室条件差异,显色时间会有所不同。此时肉眼可 见标准品的前 3-4 孔有明显的梯度蓝色,后 3-4 孔差别不明显)。
10.按每孔 0.1ml 依次加入 TMB 终止液,此时蓝色立转黄色。
11.用酶标仪在 450nm 测定 OD值。
客户购买子科生物试剂盒须知:
1、子科生物所提供的试剂产品,均为科研试剂,仅供科研使用,不得用于注射、药品、食用,血更不能用于临床检测和动物防疫检测或其他用途,对此造成的任何利润与损失、间接损失、损害承担责任经济纠纷,我司不承担任何责任。
2、买方在提货或收货时,请仔细检查货物包装,如发现包装破损,应拒收或拍照取证,随后转发给我司,买方一旦签收,我司将不对产品的包装状态负责;买方收到货品后,应及时清点,若发现货物短缺,应于签收当天通知我司,若签收次日我司尚未收到买方通知,则视为买方确认无短缺。
3、我们向您保证我们提供的产品均经过鉴定为合格产品,如果真的有任何质量问题,请您在收到产品1个月内以书面的形式及时通知我司,超过时间不予受理,我们将给您提供*周到及时的技术售后处理。人α肌动蛋白(αActin)ELISA检测试剂盒其他相关产品如下:
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Assay procedure
1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.
3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesn’t add anyting.
4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.
5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.
6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.
7. Add 50μl Stop 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.
8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.
Calculation of results
1.This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X) axis.
2.First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software.
3.To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration.
4.Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.
5.The sensitivity by this assay is 10.0 pg/ml
6.Standard curve
Storage: 2-8℃.
validity: six months.