Product Description
过氧化物酶标记试剂盒-SH 主要用于制备过氧化物酶标记的 IgG 用于酶免疫测定 (EIA) 和制备过氧化物酶标记的抗原用于竞争性 EIA。 SH 反应性过氧化物酶是该试剂盒的一个组成部分，可与蛋白质或其他分子的硫醇基团发生反应（图 1）。 该试剂盒包含标记过程所需的所有试剂，包括还原剂和储存缓冲液。 SH反应性过氧化物酶与靶分子形成共价键。 还原剂可以在 IgG 分子中产生游离硫醇基团。 当过氧化物酶标记的 IgG 用于 EIA 时，SH 反应性过氧化物酶的标记效率足够高，无需任何标记后纯化过程。如果标记后需要高纯度偶联物，只需使用亲和柱或凝胶渗透柱 . 标记小分子时，可以使用该套件中包含的过滤管去除多余的分子。
Fig. 1 IgG labeling reaction of SH-reactive peroxidase
Precaution♦ The molecular weight of the reduced protein to be labeled with this kit should be greater than 50,000.♦ The molecular weight of the small thiol compound to be labeled with this kit should be smaller than 5,000.♦ IgG or peroxidase-conjugated IgG is always on the membrane of the filtration tube during the labeling process.♦ If the IgG solution contains other proteins with molecular weight larger than 10,000, such as BSA or gelatin, purify the IgG solution prior to labeling peroxidase with this kit. IgG solution can be purified by IgG Purification Kits (not included in this kit).♦ If the IgG solution contains small insoluble materials, centrifuge the solution and use the supernatant for the labeling.
Sandwich ELISA
Fig. 2 Sandwich ELISA of human TNF-α detection
Plate: 2 μg/ml anti-human TNF-aantibody (rabbit, polyclonal)-coated high binding platerecombinant human TNF-a: 0-1000 pg/ml PBSTPeroxidase-conjugated anti-human TNF-aantibody: Prepared by Peroxidase Labeling Kit-SH.1μg/mlPBST+blocking reagentSubstrate: TMB peroxidase substrate
References1. K. Inoue, A. Sugiyama, P. C. Reid, Y, Ito, K. Miyauchi, S. Mukai, M. Sagara, K. Miyamoto, H. Satoh, I. Kohno, T. Kurata, H. Ota, A. Mantovani, T. Hamakubo, H. Daida and T. Kodama, Establishment of a High Sensitivity Plasma Assay for Human Pentraxin3 as a Marker for Unstable Angina Pectoris, Arterioscler. Thromb. Vasc. Biol., 2007, 27, 161
Yes, please follow the labeling protocol for IgG. The recovery of the conjugate should be over 80%.
Yes, if the molecular weight of the reduced form is greater than 50,000 or less than 5,000, and it has a reactive SH group, or a disulfide group that can be reduced without losing activity. If the molecular weight is greater than 50,000, follow the labeling protocol for IgG, and use 0.5-1 nmol of sample protein for LK09-10. If the molecular weight is less than 5,000, follow the labeling protocol for small molecules. If the molecular weight is between 5,000 and 50,000, contact our customer service at info@dojindo.com or 1-877-987-2667 for more information.
Yes, if the molecular weights of the oligonucleotide or the oligopeptide are less than 5,000 and they have at least one SH group. Follow the labeling protocol for small molecule.
The minimum amount is 50 μg. There is no significant difference in sensitivity and background between 50 μg and 200 μg of IgG. However, even 10 μg IgG can be labeled using 1/5 volume of SH-reactive peroxidase solution at Step 8.
The average number of peroxidase molecule per reduced IgG is 1 to 2.
If the protein solution does not contain small molecules with reactive SH groups and the concentration of the protein is 10 mg per ml, or about 70 μM, there is no need to use the Filtration tube. Just mix the sample solution with Solution B and add the mixture to a vial of the SH-reactive peroxidase.
No, you don’t have to use Storage buffer from the kit. You can choose any kind of buffer appropriate for your experiment. However, the Storage buffer helps to increase the stability of the peroxidase conjugate.
Spin the sample and use the supernatant for labeling.
No. Nearly 100% of SH reactive peroxidase is used for the IgG labeling or the small molecule labeling.
No, Storage buffer does not contain any animal products, polymers, or heavy metal ions.
Dojindo细胞分析
细胞活力和细胞毒性测定用于药物筛选和化学物质的细胞毒性测试。Dojindo开发了高度水溶性的四唑盐，称为WST。WST-8是高度稳定的WST，用于Cell Counting Kit-8（CCK-8）。由于WST-8甲maz是水溶性的，因此不会形成晶体。因此，不需要诸如MTT测定的增溶过程。此外，CCK-8的检测灵敏度高于其他四唑盐，例如MTT，XTT，MTS或WST-1。
WST检测机制
ß-半乳糖苷酶检测试剂
细胞增殖/细胞毒性转染细胞染色细胞内荧光探针细菌染色微生物活力测定干细胞分化SPiDER-ßGal线粒体检测细胞代谢
| 应用 | 产品展示 |
| 细胞生长检测，药物筛选，比色/荧光检测 | 细胞计数试剂盒-8 细胞计数试剂盒8 + 96孔有机硅定向剂 细胞计数试剂盒-F 细胞毒性LDH检测试剂盒 -WST 96孔有机硅定向剂 MTT |
| 了解检测机制的差异： | 点击这里 |
| 细胞周期分析 | 细胞周期测定溶液深红色 细胞周期测定溶液蓝色 |