This assay is intended to be conducted in a cuvette. There is no specific guidance supplied for this protocol to be used in a multi-well plate format. Please see the link below to review the full Technical Bulletin:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/228/218/sialicqbul.pdf
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运输
wet ice
储存温度
2-8°C
一般描述
唾液酸定量检测试剂盒提供了一种快速精确的测定总N-乙酰神经氨酸（NANA；唾液酸）的方法。NANA是通过神经氨酸酐酶从糖复合物中释放出来的。该试剂盒使用了α(2→3,6,8,9)神经氨酸酐酶来剪切所有的NANA连接，其中包括α(2→8)及α(2→9)连接，以及分支状NANA等，可对总唾液酸含量进行最精确的测定。
应用
含有定量自由的、或糖蛋白、细胞表面糖蛋白、聚唾液酸及荚膜多糖（仅由聚唾液酸组成）中的N-乙酰神经氨酸所需的所有试剂。
数量
试剂可用于进行25次反应，其中包括对照样品。每次反应可检测1-200 nmole的NANA。
仅试剂盒组分
产品编号
说明
- N-Acetylneuraminic acid aldolase 25 μL
- N-Acetylneuraminic acid 200 μL
- α(2→3,6,8,9)-Neuraminidase 25 μL
- Fetuin, bovine .5 mg
- Lactic dehydrogenase 25 μL
- β-NADH disodium salt 3 vial(s)
- Tris-HCl, pH 7.5 1 mL
- Sialidase buffer 500 μL
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储存分类代码
10 - Combustible liquids
其他客户在看
Corina Daniela Ene et al.
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Posttranslational modifications are dynamic enzymatic-mediated processes, regulated in time and space, associated with cancer development. We aimed to evaluate the significance of posttranslational modifications in the pathogenesis of clear cell renal cell carcinoma. The authors developed a prospective, observational study
A P Corfield et al.
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The anomeric specificity of six sialidases (Vibrio cholerae, Arthrobacter ureafaciens, Clostridium perfringens, Newcastle disease virus, fowl plague virus and influenza A2 virus sialidases) was assessed with sialylated antifreeze glycoprotein, ovine submandibular gland glycoprotein and alpha 1-acid glycoprotein, resialylated specifically in
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Analysis of glycoprotein-associated oligosaccharides.
R A Dwek et al.
Annual review of biochemistry, 62, 65-100 (1993-01-01)
商品
Understand sialic acid structure, function, signaling, and modifications. Easily find products for sialic acid research.
Understand sialic acid structure, function, signaling, and modifications. Easily find products for sialic acid research.
Understand sialic acid structure, function, signaling, and modifications. Easily find products for sialic acid research.
Understand sialic acid structure, function, signaling, and modifications. Easily find products for sialic acid research.
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How does the size of the multiwell plate affect the volumes of reagents used, and how is the final calculation of nmoles of NANA adjusted accordingly?
1 answer-
Helpful?
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How should a time course experiment be conducted to assess if digestion has reached completion? Can NANA on a glycoprotein be measured in multiple samples simultaneously using a multiwell plate as detailed in section 2, "Assay of Multiple Samples for Free NANA," or is this method only applicable to free NANA?
1 answer-
1.This kit utilizes α(2→3,6,8,9) neuraminidase to cleave all NANA linkages, including α(2→8) and α(2→9) linkages, as well as branched NANA. To determine when the digestion is complete, measurements of free NANA released at different times can be taken and plotted on a graph to illustrate the change in NANA concentration over time. The endpoint of digestion is identified as the point where no further increase in product concentration is observed, allowing for accurate determination of the time required to completely digest all the NANA in a sample.
2. For a time course experiment, one may start with an immediate baseline reading and continue with readings at 1, 2, and 3 hours to monitor the release of NANA. If the released NANA levels off between the 3 to 5-hour marks, the reaction is complete. For complex samples or high NANA concentrations, extending the incubation time points beyond 5 hours, and checking hourly, may be necessary. The optimal incubation time and/or the sample quantity would need to be adjusted based on preliminary findings to ensure complete digestion without degradation.
3.Adapting this protocol for a multiwell plate involves performing the enzymatic digestion as described in the procedure for the target sample type (e.g., free NANA, glycoproteins, or polysialic acid). After the digestion step, one would follow the steps outlined in the "2. Assay of Multiple Samples for Free NANA" section. Adjustments to the volume may be needed based on the well size of the multiwell plate. For example, for a 100 μL well, the volumes of the reagents would need to be adapted proportionally, and the final calculation of nmoles of NANA should reflect the smaller reaction volume.Helpful?
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