Pierce Mass Spec Sample Prep Kit for Cultured Cells 用于培养细胞的质谱样品制备试剂盒
货号：84840
规格：20 reactions
价格：0
产品类型：蛋白提取/纯化
品牌：Thermo Fisher
| The Thermo Scientific Pierce Mass Spec Sample Prep Kit for Cultured Cells is an easy-to-use, comprehensive kit for preparation of clean peptide mixtures from cultured cells for mass spectrometry (MS) analysis.The Pierce Mass Spec Sample Prep Kit for Cultured Cells contains all the necessary reagents and enzymes to prepare up to 20 samples (1 million cells each) for MS analysis. The simple and robust workflow uses a lysis protocol that generates approximately 100 µg of protein per sample and includes a control digestion indicator to monitor the efficiency of the two-step enzymatic digestion protocol. The kit procedure, which includes a Digestion Indicator as an internal protein/peptide control, produces consistently and reproducibly clean peptide mixtures for protein identification.Sample prep for mass spectrometry (MS, mass spec) remains one of the largest bottlenecks associated with MS analysis, and consistent and reproducible sample preparation can make the difference between a successful analysis and a failed outcome. Current sample prep protocols are primarily homebrew and can be highly variable, making data analysis and interpretation difficult. The Mass Spec Sample Prep Kit for Cultured Cells provides researchers with all the necessary tools to generate consistent and reproducible protein digests that are directly compatible with LC-MS workflows. The digests do not require further processing, such as C18 clean up or detergent removal.The included Pierce Digestion Indicator, which can also be purchased separately, is a non-mammalian recombinant protein (26kDa) with five signature peptides for use in determining the digestion efficiency and reproducibility across multiple samples. The protein sequence and recommended peptides to monitor across samples are supplied with the product instructions. |
| 特点： |
| - Complete—includes all reagents, a digestion indicator control, proteases and an optimized protocol needed to process up to 20 samples- Simple—user friendly kit can provide reproducible results even for non-expert MS analysts- Flexible—can be adapted to handle sample sizes between 10 to 200 µg- High yield—total protein yield from 1 million cells is greater than 100 µg- Optimized—cysteine reduction and alkylation are 100% with less than 1% over alkylation of non-cysteine residues- Efficient - percentage of missed cleavages is less than 10%- Compatible—final preparation is ready for direct MS analysis and other downstream applications, including mass-tag labeling |
| 数据： |
HeLa cell lysate (200µg in 200µL of Lysis Buffer and spiked with 2µg Digestion Indicator) was processed by the Thermo Scientific Pierce Mass Spec Sample Prep Kit for Culture Cells and then analyzed by mass spectrometry. Comparison of protein yields by four mass spec sample prep lysis methodsFrom one culture of HeLa S3 cells, duplicate pellets containing 2 x 10^6 cells were resuspended and lysed using 0.2mL of the respective buffers and protocol of each method (all include a sonication step); then protein concentrations and yields were determined. - FASP: 0.1M Tris-HCl, 4% SDS, 0.1M DTT, pH 7.6
- AmBic-SDS: 0.05M ammonium bicarbonate, 0.1% SDS, pH 8.0
- Urea: 0.1M Tris-HCl, 8M urea, pH 8.5
- Kit Buffer: Lysis Buffer from the Pierce Kit (Part No. 84840)
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