人CD28分子（CD28）ELISA试剂盒@2022说明书已更新《新闻》

实验原理：                                     

将目标抗体包被于微孔板中，制成固相载体，向微孔中分别加入标准品或标本，其中的目标连接于固相载体上的抗体结合，然后加入微生物化的目标抗体，将未结合的生物素抗体洗净后，加入HRP标记和亲和素，再次洗涤后加入TMB底物显色。TMB在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目标呈正相关。用酶标仪在450nm波长下测定吸光度（O.D.值），计算样品浓度。

试剂盒组成：

ELISA（酶联免疫吸附测定）是一种基于多孔板的免疫测定法，将通常作为检测成分的抗体或样品吸附在固体表面（此方法采多孔板）。ELISA以相对低廉的成本，快速、定量且灵敏地检测分析物，是简单的分析方法。此外，ELISA还可轻松改造为更高通量筛选方法，帮助研究人员通过单次运行检测大量样品。夹心法、直接法和竞争法ELISAs

我们的ELISA采用简单易用的方法，可用于研究血清、血浆、细胞培养上清液或裂解液等各种基质中的可溶性蛋白标志物。包括1000多种“仅供研究使用”的检测试剂盒，适用的样本类型包括人、小鼠、大鼠等各种动物模型（如农业和伴生动物）。每份试剂盒都提供特定实验方案及96孔板所需的试剂。ELISA试剂盒主要为96孔规格，目标物检测和定量所需试剂都由厂商提供。采用夹心法、直接法和竞争法等各种ELISA方法。

夹心法使用针对同一目标抗原不同表位的一对单克隆抗体(mAb)。固定在微孔板中的一抗，用于将蛋白从溶液中提取出来。二抗则用于完成“夹心”，提供表示目标抗原存在的信号。试剂盒提供重组形式的已知含量的抗原蛋白，方便用户建立标准曲线解析样品信号。我们大部分产品采用的都是这种夹心法ELISA。

直接法和竞争法比较少见。直接法先用一种单抗检测固定在微孔板上的样品。然后一抗会与的酶标二抗结合提供信号。

竞争法在微孔板上预包被已知含量的生物标志物。酶标抗体与样品共同孵育，并与预包被抗体进行“竞争性”结合反应，具体结果取决于样品中目标物的浓度。于是游离抗体能与微孔板上的抗原结合，并在样品洗去后呈现信号。该信号与目标标志物的浓度成反比。

血清：用无菌管收集，室温血液自然凝固10-20分钟，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如出现沉淀，应再次离心。

 

2、血浆：应根据标本的要求选择EDTA或柠檬酸钠作为抗凝剂抗凝剂，混合10-20分钟后，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如有沉淀形成，应该再次离心。

3、尿液：用无菌管收集，2-8℃条件离心20分钟左右（2000-3000转/分）。仔细收集上清，保存过程中如有沉淀形成，应再次离心。

4、胸腹水：用无菌管收集，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如有沉淀形成，应再次离心。

5、脑脊液：用无菌管收集，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如有沉淀形成，应再次离心。

6、上清，保存过程中如有沉淀形成，应再次离心。

7、细胞培养上清：检测分唾液：用无菌管收集，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集泌性的成份时，用无菌管收集。2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如有沉淀形成，应再次离心。

8、牛奶：用无菌管收集，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如有沉淀形成，应再次离心。

9、蜂蜜：用无菌管收集，2-8℃条件离心20分钟左右（2000-3000转/分），仔细收集上清，保存过程中如有沉淀形成，应再次离心。

10、全血：用含有抗凝剂的无菌管收集，立即轻轻摇动，来回轻轻颠倒数次，使血液和抗凝剂混匀，以防血液凝固。

Experiment principle:

The target antibody is coated in the microplate to form a solid phase carrier. The standard or sample is added to the microplate respectively, where the target is connected to the antibody on the solid phase carrier for binding, ａnd then the microbial target antibody is added. After the unconjugated biotin antibody is washed, HRP labeling ａnd idin are added, ａnd TMB substrate is added for color development after thorough washing again. TMB is converted to blue under the catalysis of peroxidase, ａnd finally to yellow under the action of acid. The depth of color is positively correlated with the target in the sample. Use the microplate reader to measure the absorbance (O.D. value) at 450nm welength ａnd calculate the sample concentration.

Kit composition:

ELISA (enzyme-linked immunosorbent assay) is an immunoassay based on a porous plate. Antibodies or samples that are usually used as one of the test components are adsorbed on the solid surface (porous plate is used for this method). ELISA is one of the simplest analytical methods with relatively low cost, rapid, quantitative ａnd sensitive detection of analytes. In addition, ELISA can be easily transformed into a higher flux screening method to help researchers detect a large number of samples through a single run. Sandwich method, direct method ａnd competition law ELISAs

 

Our ELISA is a simple ａnd easy to use method, which can be used to study soluble protein markers in various matrices such as serum, plasma, cell culture supernatant or lysate. It includes more than 1000 "only for research" test kits, ａnd the applicable sample types include human, mouse, rat ａnd other animal models (such as agriculture ａnd companion animals). Each kit provides the reagents required for a specific experimental protocol ａnd 96 well plate. The ELISA kit is mainly 96 hole specification, ａnd the reagents required for target detection ａnd quantification are provided by the manufacturer. ELISA methods such as sandwich method, direct method ａnd competitive method were used.

The sandwich method uses a pair of monoclonal antibodies (mAb) targeting different epies of the same target antigen. The first antibody fixed in the microplate is used to extract the protein from the solution. The second antibody is used to complete the "sandwich" ａnd provide a signal indicating the presence of the target antigen. The kit provides recombinant antigen protein with known content, which is convenient for users to establish standard curves to analyze sample signals. Most of our products use this sandwich ELISA.

Direct law ａnd competition law are rare. The direct method first uses a monoclonal antibody to detect the sample fixed on the microplate. The first antibody then binds to the enzyme labeled second antibody to provide a signal.

Biomarkers of known content are pre coated on the microplate by competitive method. The enzyme labeled antibody is co incubated with the sample ａnd "competitively" reacts with the pre coated antibody. The specific result depends on the concentration of the target in the sample. Thus, the free antibody can bind to the antigen on the microplate ａnd present a signal after the sample is washed away. This signal is inversely proportional to the concentration of the target marker.

Serum: collected with sterile tube, the blood naturally coagulates at room temperature for 10-20 minutes, centrifuges at 2-8 ℃ for about 20 minutes (2000-3000 rpm), carefully collects the supernatant, ａnd centrifuges again if precipitation occurs during storage.

2. Plasma: EDTA or sodium citrate should be selected as the anticoagulant ａnd anticoagulant according to the requirements of the sample. After 10-20 minutes of mixing, centrifugate at 2-8 ℃ for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant. If precipitation forms during storage, centrifugate again.

3. Urine: collected with sterile tube, centrifuged at 2-8 ℃ for about 20 minutes (2000-3000 rpm). Carefully collect the supernatant, ａnd centrifugate again if sediment is formed during storage.

4. Thorax ａnd ascites: collect with sterile tube, centrifugate at 2-8 ℃ for about 20 minutes (2000-3000 rpm), carefully collect the supernatant, ａnd centrifugate again in case of precipitation formation during storage.

5. Cerebrospinal fluid: collected with sterile tubes, centrifuged at 2-8 ℃ for about 20 minutes (2000-3000 rpm), carefully collected the supernatant, ａnd centrifuged again if sediment is formed during storage.

6. The supernatant shall be centrifuged again in case of precipitation formation during storage.

7. Cell culture supernatant: detect saliva: collect with sterile tube, centrifugate at 2-8 ℃ for about 20 minutes (2000-3000 rpm), ａnd carefully collect the secreted ingredients with sterile tube. Centrifuge at 2-8 ℃ for about 20 minutes (2000-3000 rpm), carefully collect the supernatant, ａnd centrifugate again if sediment is formed during storage.

8. Milk: Collect with sterile tube, centrifugate at 2-8 ℃ for about 20 minutes (2000-3000 rpm), carefully collect the supernatant, ａnd centrifugate again if sediment is formed during storage.

9. Honey: Collect it with a sterile tube, centrifuge at 2-8 ℃ for about 20 minutes (2000-3000 rpm), carefully collect the supernatant, ａnd centrifuge again if sediment forms during storage.

10. Whole blood: Collect it with a sterile tube containing anticoagulant, shake it gently immediately ａnd turn it back ａnd forth slightly for several times to mix the blood ａnd anticoagulant to prevent blood coagulation.

牛血管内皮生长因子B（VEGFB）ELISA试剂盒@2022说明书已更新《新闻》