High-Select™ 磷酸化多肽富集试剂盒和试剂
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High-Select™ 磷酸化多肽富集试剂盒和试剂
Thermo Scientific™

High-Select™ 磷酸化多肽富集试剂盒和试剂

经扩展的 High Select 系列提供了完备的试剂盒,支持使用离心柱和吸头中的 Fe-NTA 和 TiO2 固定化树脂以及磁珠(专为 MS 分析而设计)富集磷酸化肽。
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货号数量产品类型
A3299230 次反应Fe-NTA磷酸化多肽富集试剂盒
A3299324 次反应TiO2磷酸化多肽富集试剂盒
8830230柱色谱柱
8830396 吸头TiO2 磷酸多肽富集离心吸头
货号 A32992
价格(CNY)
4,021.00
飞享价
Ends: 31-Dec-2025
5,208.00
共减 1,187.00 (23%)
Each
添加至购物车
数量:
30 次反应
产品类型:
Fe-NTA磷酸化多肽富集试剂盒
价格(CNY)
4,021.00
飞享价
Ends: 31-Dec-2025
5,208.00
共减 1,187.00 (23%)
Each
添加至购物车
质谱分析 (MS) 是鉴别蛋白磷酸化位点和定量磷酸化变化的关键工具。然而,由于磷酸肽的化学计量值较低、亲水性较高、电离不理想且未完全片段化,因此蛋白磷酸化的 MS 分析极具挑战。鉴于复合蛋白样品中磷酸化修饰的相对丰度较低,富集对于成功进行磷酸肽 MS 分析至关重要。High-Select Fe-NTA 和 TiO2 磷酸肽富集试剂盒补充了我们的裂解、还原、烷基化和消化试剂,以及我们的 C18、石墨离心和高 pH 值反相肽分馏柱,从而提供完整的磷酸肽富集工作流程。
便利—以离心柱(Fe-NTA 和 TiO2)或磁珠 (Fe-NTA) 形式提供预配制缓冲液,易于使用,能够对多份样品进行平行处理
高特异性—以 > 90% 的选择性回收磷酸肽
出色的回收率—与其他市售树脂相比,能够富集较多总磷酸肽和独特磷酸肽
高覆盖范围—回收具有单一和多磷酸化位点的肽
可扩展—兼容自动化操作,包含使用 KingFisher 磁珠纯化仪

High-Select Fe-NTA 磷酸肽富集试剂盒使用离心柱中的铁螯合树脂从复杂样品中富集磷酸肽。每柱可富集 0.5 至 5 mg 总蛋白酶切物的磷酸肽,作为起始样品。配合使用预配制缓冲液,改进和简化的程序可在不到 45 分钟内富集多达 150 µg 的磷酸肽,选择性大于 90%。

High-Select TiO2 磷酸肽富集试剂盒包括 24 个二氧化钛 (TiO2) 离心吸头和优化的缓冲液,可促进富集磷酸肽的制备。每柱可富集 0.5 至 3 mg 总蛋白酶切物的磷酸肽,作为起始样品。经优化的缓冲液和简化的程序消除了对洗脱缓冲液中的吡咯烷和使用石墨离心柱进行后续清理的需求。使用该试剂盒可以富集多达 150 µg 的磷酸肽,选择性大于 90%。TiO2 富集了一组独特的磷酸肽,可补充 High-Select Fe-NTA IMAC 磷酸肽富集试剂盒。

High-Select Fe-NTA 磁性磷酸肽富集试剂盒使用铁螯合磁性琼脂糖微珠,从复杂样品中富集磷酸肽。每次反应足以在不到 45 分钟内从 0.5–5 mg 总蛋白酶切物富集 250–1000 µg 磷酸肽,作为起始样品。此外,试剂盒也可用于富集含官能团的化学修饰肽,设计用于配合固定化金属亲和色谱树脂一起使用。

High Select Fe-NTA 磁性琼脂糖专门设计用于富集磷酸肽和化学修饰肽,以进行 MS 分析。我们的非聚集性磁石 (Fe3O4) 是一种超顺磁性微珠,具有出色的均匀性,可在手动和自动纯化应用中实现高性能纯化。Fe-NTA 磁性琼脂糖可单独购买,满足实验设计的灵活性。

对于富集磷酸肽的高分辨率分析,推荐用于 Nanospray Flex 源的 LC 色谱柱为 Acclaim PepMap 100 C18 LC 色谱柱(货号 164942 或 164939)。对于 EASY-Spray 离子源,推荐的 LC 色谱柱为 EASY-Spray C18 LC 色谱柱(货号 ES902 或 ES903)。独立的 High Select Fe-NTA Magnetic Agarose 微珠可灵活设置规模和通量,可用于手动和自动化平台,包含 KingFisher 96 和 KingFisher Flex 磁粒子处理器。

For Research Use Only. Not for use in diagnostic procedures.
规格
最终产品类型
适用于(设备)小型离心机
标签或染料未标记
数量30 次反应
运输条件湿冰
工作流程步骤肽富集
检测方法质谱分析
产品规格试剂盒
产品线High Select™
产品类型Fe-NTA磷酸化多肽富集试剂盒
原始材料蛋白酶酶切的蛋白
Unit SizeEach
内容与储存
• 离心柱 (30)
• 白色鲁尔插塞(端盖)(30)
• 结合/洗涤缓冲液 (2 x 20 mL)
• 磷酸肽洗脱缓冲液 (7 mL)

常见问题解答 (FAQ)

Should I desalt my peptide samples before enrichment using the High Select Phosphopeptide Enrichment Kits?

Yes, we do recommend desalting peptide samples before enrichment when using High-Select Fe-NTA and TiO2 phosphopeptide enrichment kits. This helps ensure optimal results by removing detergents and salts that can interfere with the enrichment process.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

In the SMOAC protocol (https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/10/High-SelectTM-SMOAC-Protocol.pdf), can I enrich with High-Select Fe-NTA kit first?

No. It is important to enrich with the TiO2 kit (Cat. No. A32993) first. Afterwards, the flow-through and wash fractions from this enrichment can be processed with the Fe-NTA kit (Cat. No. A32992). If this order is reversed (that is, Fe-NTA before TiO2), there will be 2 consequences as follows: 1. There will not be any significant additional recovery of peptides (maybe just a few more peptides). 2. There will be no enrichment for the multiple phosphorylated peptides, so those would be lost.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Why do you offer two phosphopeptide enrichment kits: the High-Select TiO2 kit (Cat. No. A32993) and the High-Select Fe-NTA kit (Cat. No. A32993)?

The two phosphopeptides enrichment kits, Fe-NTA and TiO2, enrich a complementary set of phosphopeptides. Our R&D has developed a Sequential enrichment of Metal Oxide Affinity Chromatography (see https://assets.thermofisher.com/TFS-Assets/CMD/posters/PO-65032-SMOAC-Phosphoproteomics-Peptides-ASMS2017-PO65032-EN.pdf and https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/10/High-SelectTM-SMOAC-Protocol.pdf?CID=bid_mic_r04_jp_cp0000_pjt0000_bid00000_0so_blg_protein_analysis_mass_spectrometry_bid_ts_mbr_24065_Social_LAB) where flow-through and wash fractions from TiO2 enrichment were combined and subjected to Fe-NTA enrichment. This sequential enrichment provides impressive coverage of phosphoproteomes.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What are the differences between the old Fe-NTA kit (Cat. No. 88300) and the new High-Select Fe-NTA kit (Cat. No. A32992)?

There are four differences between the Fe-NTA kit (Cat. No. 88300) and the new High-Select Fe-NTA kit (Cat. No. A32992) kit as follows: 1. The selectivity - the ratio of number of phosphopeptides over total peptides - was significantly improved to 99% with Cat. No. A32992, because the reagents were extensively optimized for the phosphopeptide selection. 2. The phosphopeptide yield was also increased to 33 µg based on quantitative colorimetric peptide assay (Cat. No. 23275). 3. The reagent is a pre-formulated format, so mixing reagent to prepare the working solution from stock solutions provided in the old kit (Cat. No. 88300) is not necessary, so it is easier to handle. 4. The enrichment protocol is optimized and streamlined, which means there are many fewer steps than with Cat. No. 88300. Thus, it takes <45 min to finish the entire protocol compared to 2 hours with the old kit (Cat. No. 88300).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What type of samples can I use with the High-Select Fe-NTA Phosphopeptide Enrichment Kit and High-Select Fe-NTA Magnetic Phosphopeptide Enrichment Kit?

The High-Select Fe-NTA Phosphopeptide Enrichment Kit and High-Select Fe-NTA Magnetic Phosphopeptide Enrichment Kit are compatible with desalted and detergent-free protein digests or lyophilized peptide fractions.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.