In proteomics research, precise protein digestion is a critical step to ensure the accuracy and coverage of mass spectrometry analysis. Endoproteinase rGlu-C (MS grade) is a serine protease that specifically cleaves peptide bonds at the C-terminal side of glutamic acid residues. In ammonium bicarbonate or ammonium acetate buffers, the enzyme exhibits high specificity for glutamic acid. In phosphate buffer, it can cleave both aspartic acid and glutamic acid residues. The optimal pH range for rGlu-C activity is 4.0-9.0. This MS-grade enzyme can be used alone or in combination with other proteases to generate diverse proteolytic peptides from protein substrates.
Product Overview
The sequencing grade rGlu-C provided by MtoZ Biolabs is a high-purity recombinant enzyme suitable for various protein digestion applications, exhibiting excellent stability and enzymatic activity within a pH range of 4.0-9.0, making it particularly effective for in-solution digestion protocols. It enables researchers to achieve more comprehensive and accurate proteomic data. rGlu-C can be used alone or in combination with trypsin to enhance peptide coverage and improve mass spectrometry analysis outcomes.
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Product Size |
rGlu-C Lyophilized Powder (50 μg) |
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Molecular Weight |
24 kDa |
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Storage Condition |
Lyophilized powder at -20℃ Reconstituted solution at -20℃ for up to 8 weeks |
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Optimal pH |
4.0-9.0 |
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Shelf Life |
36 months at -20℃ |
Protocol
1. Enzyme Reconstitution and Storage
(1) Reconstitute using 50 mM ammonium bicarbonate or phosphate buffer (pH 7.8).
(2) Store long-term at -20 °C or -80 °C; avoid repeated freeze-thaw cycles.
2. Recommended Digestion Conditions
(1) Applicable pH range: 4.0-9.0
(2) Recommended enzyme-to-substrate ratio: 1:20 (w/w)
(3) Temperature: 37°C, typically 4-16 hours (adjust based on experimental requirements)
3. Digestion Termination Methods
To terminate digestion and prevent over-digestion, the following methods can be used:
(1) Lower the temperature: Store the sample at -20°C or lower.
(2) Add termination reagents: For example, add trifluoroacetic acid (TFA) to a final concentration of 0.1%.
Features and Benefits
1. High Cleavage Specificity
Enables targeted cleavage at glutamic acid or aspartic acid residues depending on the buffer system, enhancing sequence coverage.
2. High Purity with Low Background
Produced via tightly controlled recombinant expression, free from contaminating proteins, minimizing background interference and improving data quality.
3. Excellent Stability
Maintains enzymatic activity across a broad pH range, suitable for diverse experimental designs.
4. Strong Buffer Compatibility
Compatible with a variety of buffer systems (e.g., ammonium bicarbonate, ammonium acetate, phosphate), offering greater experimental flexibility.
5. Multi-Platform Adaptability
Compatible with a wide range of mass spectrometry platforms including MALDI-TOF, Q-TOF, and Orbitrap, applicable to various biological sample types.
Applications
1. Proteomics Research
Provides a cleavage pattern complementary to trypsin, enhancing mass spectrometry identification accuracy and protein sequence coverage.
2. Protein Sequencing Analysis
Precisely cleaves long peptides to aid in sequence assembly and full-length protein validation.
3. Post-Translational Modification (PTM) Analysis
Improves the detection of modification sites such as phosphorylation and glycosylation, increasing PTM analysis sensitivity.
4. Structural Biology Research
Accurately defines domains and functional sites, supporting studies of protein conformation and function.
5. Complex Protein Mixture Analysis
Efficiently digests highly complex samples, increasing peptide detectability and enhancing mass spectrometry performance.
MtoZ Biolabs' sequencing grade rGlu-C offers high specificity, excellent stability, and broad compatibility, providing an efficient and precise solution for proteomics and protein analysis. It empowers researchers to conduct in-depth and accurate exploration of protein structure and function.
FAQs
Q1: What Is the Primary Cleavage Site of rGlu-C?
A1: rGlu-C specifically cleaves at the C-terminal side of glutamic acid (E) or aspartic acid (D) residues, with specificity depending on the buffer system used.
Q2: How Does rGlu-C’s Cleavage Specificity Vary between Different Buffer Systems?
A2: In ammonium bicarbonate and ammonium acetate buffers, rGlu-C predominantly cleaves at glutamic acid (E) residues. In phosphate buffer, cleavage extends to both glutamic acid (E) and aspartic acid (D) residues.
Q3: Can rGlu-C Be Used for Phosphorylation or Other Post-translational Modification (PTM) Studies?
A3: Yes, peptides generated by rGlu-C digestion are well-suited for capturing information on phosphorylation, glycosylation, and other PTMs.
