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  • ‹ Back to Mass spectrometry reagents
  • Capturem trypsin digestion for mass spectrometry
  • Capturem pepsin digestion
  • Trypsin digestion—TPCK magnetic trypsin
Technical notes View data: Capturem Pepsin protein digestion
Home › Products › Protein research › Mass spectrometry reagents › Capturem pepsin digestion

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Technical notes View data: Capturem Pepsin protein digestion

Capturem Pepsin digestion for mass spectrometry (Phase-out notice: These products will not be available after current inventory is exhausted)

Pepsin mini spin columns for rapid, complete protein digestion for mass spectrometry

Capturem Pepsin provides fast, efficient, and complete digestion of proteins at room temperature, in comparison to typical protein digestion workflows for mass spectrometry (MS) analyses that require four-hour or overnight digestion. By contrast, Capturem Pepsin's rapid and simple protocol allows for an uninterrupted proteomic analysis workflow. This product, based on our novel Capturem technology, provides membrane-immobilized pepsin in a mini spin column format.

Capturem Pepsin provides fast, efficient, and complete digestion of proteins at room temperature, in comparison to typical protein digestion workflows for mass spectrometry (MS) analyses that require four-hour or overnight digestion. By contrast, Capturem Pepsin's rapid and simple protocol allows for an uninterrupted proteomic analysis workflow. This product, based on our novel Capturem technology, provides membrane-immobilized pepsin in a mini spin column format. Capturem Pepsin completely digests protein samples in just a few minutes, compared to the time-consuming incubations often required for in-solution digestion, and provides better efficiency with fewer autolyzed fragments. In addition, this Capturem spin column makes it easier to avoid the over-digestion commonly seen with in-solution digests.

More effective proteomic analysis

Protein digestion using Capturem Pepsin is more effective than in-solution digestion for preparing samples for MS analysis because it eliminates long digestion times, which may lead to oxidation and other protein modifications, as well as increased autolysis, which can make peptide and protein identification more difficult and less reproducible.

Fast, easy protocol

Capturem Pepsin includes mini spin columns and an activation buffer. Capturem protein digestion is performed using a fast, easy protocol. Just load 200 µl of activation buffer, spin for one minute, then load your protein sample and spin for another minute to elute peptides that are ready for downstream analysis.

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Cat. # Product Size License Quantity Details
635728 Capturem™ Pepsin Miniprep Kit 20 Rxns USD $271.00

License Statement

ID Number  
273 This product is covered by U.S. Patent Nos. 9,459,188 and/or 10,207,229, exclusively licensed to Takara Bio USA, Inc.

Capturem Pepsin provides fast, efficient, and complete digestion of proteins at room temperature, allowing for an uninterrupted proteomic analysis workflow. This product is based on our novel Capturem technology and provides membrane-immobilized pepsin in a mini spin column format. Capturem Pepsin completely digests protein samples in just a few minutes while avoiding over-digestion by controlling the time the sample is exposed to the protease. This kit contains Capturem Pepsin mini spin columns and an activation buffer. Each column can process 10–50 µg of sample in a loading volume of up to 850 µl, and requires a minimum elution volume of 200 µl.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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Capturem Pepsin workflow

Capturem Pepsin workflow

Capturem Pepsin workflow. Antibody samples are denatured and reduced before enzymatic digestion. After initial activation of the columns with the included Activation Buffer, the sample solution (50–800 µl) containing the reduced antibody is passed through the spin column via a one-minute centrifugation. Digestion by pepsin occurs on the column, and subsequent peptide fragments are eluted in a second one-minute centrifugation. Eluted peptides are neutralized with the addition of NaOH and ready for downstream analysis.

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Digestion of different mouse antibody isotypes with Capturem Pepsin

Digestion of different mouse antibody isotypes with Capturem Pepsin

Digestion of different mouse antibody isotypes with Capturem Pepsin. 100 µg each of mouse IgGs (IgG1, IgG2a, IgG2b, and IgG3) was denatured in TCEP and acetic acid at 75°C for 15 min, and then diluted in 5% formic acid buffer. The diluted samples were digested using Capturem Pepsin spin columns. Both digested and undigested samples (4 µg each) were evaluated by SDS-PAGE.

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HPLC profiles of lot-to-lot comparisons of Capturem Pepsin digests

HPLC profiles of lot-to-lot comparisons of Capturem Pepsin digests

HPLC profiles of lot-to-lot comparisons of Capturem Pepsin digests. 50 µg of Apo was diluted in 5% formic acid and passed through Capturem Pepsin columns from three different lots. HPLC profiles show the same fragments for each sample digestion.

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Comparison of digestion with Capturem Pepsin columns versus standard in-solution pepsin methods

Comparison of digestion with Capturem Pepsin columns versus standard in-solution pepsin methods

Comparison of digestion with Capturem Pepsin columns versus standard in-solution pepsin methods. 50 µg of anti-HER2 mAB was digested using three different methods: Capturem Pepsin, in-solution pepsin incubated for four hours, and in-solution pepsin incubated overnight. MS analysis was performed following digestion. The left shift of the peaks for the in-solution digests and the higher number of peaks overall is indicative of over-digestion. Neither of these results were observed for the Capturem Pepsin digest.

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Load volumes and approximate yields for available Capturem formats by chemistry

Load volumes and approximate yields for available Capturem formats by chemistry

Load volumes and yields of Capturem purification formats.

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Capturem spin columns and filtration devices

Capturem spin columns and filtration devices

Available Capturem spin column and filtration device formats. Pictured from left to right: nanoprep, miniprep, 96-well, 24-well, maxiprep, and large-volume Capturem formats. Formats available vary for the different functionalized membranes. Check product pages for list of available formats.

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635728: Capturem Pepsin Miniprep Kit

635728: Capturem Pepsin Miniprep Kit

Overview

  • Digests protein samples in less than two minutes instead of overnight
  • Generates fewer autolyzed pepsin fragments and protein modifications
  • Allows an uninterrupted proteomics analysis workflow
  • Enables more effective peptide and protein identification in MS analysis

More Information

Applications

  • Protein characterization
  • Proteomic analysis
  • Mass spectrometry sample preparation
  • Antibody characterization

Additional product information

Please see the product′s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


Capturem Pepsin and Capturem Trypsin

Get the data on fast, efficient protein digestion with Capturem Trypsin and Capturem Pepsin

Capturem Pepsin and Capturem Trypsin were developed for complete digestion of proteins without the need for the long incubation times typically required for in-solution protocols. Both of these kits are powerful tools that allow for an uninterrupted proteomic analysis workflow. The performance of both kits has been demonstrated using apomyoglobin, a common protein mapping standard. Apomyoglobin peptides generated by Capturem Trypsin and Capturem Pepsin digestion were shown to provide good sequence coverage without over-digestion of the samples. Click the buttons below to view technical data on each digestion kit.

Capturem Pepsin Capturem Trypsin

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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