标签：碱性磷酸酶标记试剂盒-SH

Dojindo,Alkaline Phosphatase Labeling Kit-SH/3/LK13,碱性磷酸酶标记试剂盒-SH

碱性磷酸酶标记试剂盒-SH 主要用于制备碱性磷酸酶标记的 IgG 用于酶免疫测定 (EIA) 和制备碱性磷酸酶标记的抗原用于竞争性 EIA。 SH-reactive ALP 是该试剂盒的一个组成部分，可以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 ALP 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。 当碱性磷酸酶标记的 IgG 用于 EIA 时，SH 反应性 ALP 的标记效率足以消除标记后的任何纯化过程。 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即可。 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。

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 amine compound to be labeled with this kit should be smaller than 5,000.♦ IgG or alkaline phosphatase-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 alkaline phosphatase 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.

Peroxidase Labeling Kit-NH2-used for Viral Research (COVID-19 Testing)

Dr. Kyosei et al. developed the ultrasensitive detection of spike proteins of SARS-CoV-2. The SARS-CoV-2 spike proteins were measured by a sandwich ELISA coupled with thio-NAD cycling using alkaline phosphatase, androsterone derivatives, and 3α-hydroxysteroid dehydrogenase (3α-HSD) and its coenzymes (NADH and thio-NAD). The secondary antibody was conjugated to alkaline phosphatase by using Dojindo’s Alkaline Phosphate Labeling Kit-SH.

Y. Kyosei et al., Diagnostics, 2020, 10(8), 594.“Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2”

Sandwich ELISA

Fig. 2 Sandwich ELISA of human TNF-α detection

Plate: 2μg/ml anti-human TNF-antibody (rabbit, polyclonal)-coatedhigh binding platerecombinant human TNF-a: 0-1000 pg/ml PBSTALP-conjugated anti-human TNF-antibody: Prepared by Alkaline phosphatase Labeling Kit-SH .1μg/ml PBST+blocking reagentSubstrate: p-Nitrophenylphosphate, ALP substrate

Can I use this kit for F(ab E2?
Yes, please follow the labeling protocol for IgG. The recovery of the conjugate should be over 80%.
Can I use this kit for other proteins or peptides?
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 LK13-10. 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.
Can I use this kit to label oligopeptides or oligonucleotides?
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.
What is the minimum amount of IgG that can be labeled with LK13-10?
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 ALP solution at step 8.
How many alkaline phosphatase molecules per reduced IgG are introduced?
The average number of alkaline phosphatase molecule per reduced IgG is 1 to 2.
Do I have to use a Filtration tube prior to labeling the protein?
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 10 μl of the sample solution with Solution B and add the mixture to a vial of the SH-reactive peroxidase.
Do I have to use Storage buffer included with the kit?
No, you don’t have to use Storage buffer from the kit. You can choose any kind of buffer appropriate for your experiment.
My sample contains small insoluble material. What should I do?
Spin the sample and use the supernatant for labeling.
Does unconjugated SH-reactive ALP still have a reactive maleimide after the labeling reaction to IgG?
No. Nearly 100% of SH-reactive ALP is used for the IgG labeling or the small molecule labeling.
Does Storage buffer contain animal products or polymers?
No, Storage buffer does not contain any animal products , polymers, or heavy metal ions.