サンプル調製は、いまだに質量分析法（MS）による解析に関連する最大のボトルネックの１つです。矛盾のない再現性のあるサンプルの調製は、解析が成功するか、誤った結果を導くかの別れめとなります。Thermo Scientific™ の試薬とキットは、複雑性を軽減する除去戦略から微量タンパク質の分離ためのタンパク質抽出、濃縮およびクリーンアップまでの工程の各ステップにおけるサンプル調製のために最適化されています。消化後、ペプチド濃縮、分画およびクリーンアップのステップは、適切な解析にきわめて重要となる場合が多くあります。比色定量的および蛍光定量的なペプチドアッセイは、消化後のサンプルの標準化を可能にします。
包括的 —タンパク質の消化前および消化後のサンプル調製のワークフローにおける各ステップのための試薬およびキット最適化 —すべての試薬およびキットは、アバンダントタンパク質を最大限に除去し、タンパク質とペプチドの収率を向上させるように設計されています。広範囲の用途 —一式のキットでも単独の試薬としても使用できる製品。適合性 —すべてのキットは、質量分析と適合するように設計されています。バリデート済み —すべての製品は十分に試験され、Thermo Scientific™の質量分析計を使用して分析されています。
お客さまのサンプルのタイプに合った質量分析用のサンプル調製のために最適な製品をお選びください。
質量分析用のサンプルの調製のための製品
アルブミン、存在量の多さがTop 2またはTop12のタンパク質を除去するための製品です。最も多く存在するタンパク質を除去することにより、消化に先立って、存在量の少ないタンパク質の検出を向上させます。
Thermo Scientific™ のPierce™ Mass Spec Sample Prep Kit for Cultured Cellsは、質量分析法（MS）による分析用の培養細胞から得たクリーンなペプチド混合液の調製のための使いやすい、総合的なキットです。
プロテアーゼ・フォスファターゼ阻害剤カクテルおよび錠剤タイプのプロテアーゼ・ホスファターゼ阻害剤は、タンパク質消化前に、組織および初代哺乳類細胞または培養哺乳類細胞から得たライセートを調製する際のタンパク質保護に最適です。
濃縮の手法には、免疫沈降法やPTM（リン酸化タンパク質濃縮など）や活性部位の標識による濃縮と活性依存型プローブを用いた濃縮などがあります。
シンプルかつ効率的に脱塩、バッファー交換、およびサンプルからの界面活性剤の除去を行う豊富な特殊デバイスとレジン。
質量分析法を使用した解析のために、対象のタンパク質をより小さなフラグメントまたはペプチドに分解するプロテアーゼ
存在量の少ないタンパク質の分析と翻訳後修飾されたペプチドの同定を成功させるためには、複数のステップ、すなわち濃縮、分画および／またはクリーンアップが必要です。
各処理ステップにおけるタンパク質やペプチドの濃度を標準化するのに役立つ、BCAタンパク質定量アッセイや新規のペプチド定量アッセイなどのタンパク質アッセイ
The new colorimetric and fluorescent peptide quantitation assays help to normalize the peptide concentration of samples during each processing step following protein digestion.
質量分析のためのサンプル調製についてのビデオ
Prepare clean peptide mixtures from cultured cells for mass spectrometry (MS) analysis with this easy-to-use, comprehensive kit. Sample preparation is critical to effectively identify proteins by mass spectrometry, and the Thermo Scientific Pierce Mass Spec Sample Prep Kit for Cultured Cells helps you to easily extract and digest proteins from cells for shotgun proteomics experiments. The first step in the workflow is to lyse the cells. After harvesting and washing at least 10^6 cells, suspend the cell pellet in Cell Lysis Buffer and incubate for 5 minutes at 95°C to lyse the cells. Cool the lysate on ice for 5 minutes and then sonicate it on ice until the DNA in the lysate is completely sheared, which you can tell by the loss of viscosity caused by intact genomic DNA. Separate the proteins from the DNA and cell debris by centrifugation, collect the supernatant and determine its protein concentration. The next step in the workflow is to reduce, alkylate and precipitate the proteins. Transfer 100µg protein to a new tube and adjust the volume to 100µL using the Cell Lysis Buffer. If desired, also add 0.5µg Thermo Scientific Pierce Digestion Indicator to the sample. Add 2.1µL dithiothreitol to the sample and incubate for 45 minutes at 50°C to reduce the proteins in the sample. After cooling, add 11.5µL iodoacetamide to the protein extract and incubate for 20 minutes in the dark at room temperature to alkylate the proteins in the sample. Immediately add 460µL pre-chilled 100% acetone to the sample, vortex the tube and incubate it for 1 hour to overnight at -20°C to precipitate the proteins in the sample. Pellet the proteins by centrifugation, aspirate the acetone, and then add 50µL pre-chilled 90% acetone and vortex the tube to wash the proteins. Then pellet the proteins by centrifugation, remove the acetone and dry the pellet for 2-3 minutes. The next step is to digest the proteins. First, add 100µL Digestion buffer and suspend the pellet by pipetting up and down repeatedly. Add 1µg Lys-C protease, incubate for 2 hours at 37°C, and then add 2µg trypsin protease and incubate overnight at 37°C to digest the proteins in the sample. Quench the digestion by freezing the sample at -80°C or acidifying the sample with 0.1% trifluoroacetic acid, and dry down the peptides. Then resuspend the peptides in 0.1% TFA or formic acid, and they are ready for analysis by mass spectrometry.
Prepare clean peptide mixtures from cultured cells for mass spectrometry (MS) analysis with this easy-to-use, comprehensive kit. Sample preparation is critical to effectively identify proteins by mass spectrometry, and the Thermo Scientific Pierce Mass Spec Sample Prep Kit for Cultured Cells helps you to easily extract and digest proteins from cells for shotgun proteomics experiments. The first step in the workflow is to lyse the cells. After harvesting and washing at least 10^6 cells, suspend the cell pellet in Cell Lysis Buffer and incubate for 5 minutes at 95°C to lyse the cells. Cool the lysate on ice for 5 minutes and then sonicate it on ice until the DNA in the lysate is completely sheared, which you can tell by the loss of viscosity caused by intact genomic DNA. Separate the proteins from the DNA and cell debris by centrifugation, collect the supernatant and determine its protein concentration. The next step in the workflow is to reduce, alkylate and precipitate the proteins. Transfer 100µg protein to a new tube and adjust the volume to 100µL using the Cell Lysis Buffer. If desired, also add 0.5µg Thermo Scientific Pierce Digestion Indicator to the sample. Add 2.1µL dithiothreitol to the sample and incubate for 45 minutes at 50°C to reduce the proteins in the sample. After cooling, add 11.5µL iodoacetamide to the protein extract and incubate for 20 minutes in the dark at room temperature to alkylate the proteins in the sample. Immediately add 460µL pre-chilled 100% acetone to the sample, vortex the tube and incubate it for 1 hour to overnight at -20°C to precipitate the proteins in the sample. Pellet the proteins by centrifugation, aspirate the acetone, and then add 50µL pre-chilled 90% acetone and vortex the tube to wash the proteins. Then pellet the proteins by centrifugation, remove the acetone and dry the pellet for 2-3 minutes. The next step is to digest the proteins. First, add 100µL Digestion buffer and suspend the pellet by pipetting up and down repeatedly. Add 1µg Lys-C protease, incubate for 2 hours at 37°C, and then add 2µg trypsin protease and incubate overnight at 37°C to digest the proteins in the sample. Quench the digestion by freezing the sample at -80°C or acidifying the sample with 0.1% trifluoroacetic acid, and dry down the peptides. Then resuspend the peptides in 0.1% TFA or formic acid, and they are ready for analysis by mass spectrometry.
85160,85161,85162,85164,85165,24600,24615,24594
