Plasma, as one of the most commonly studied biological fluids, is widely used in disease diagnosis, prognosis evaluation, and drug development. It contains hundreds of proteins, including high-abundance components such as albumin and immunoglobulins, as well as low-abundance proteins like hormones and enzymes. These proteins play critical roles in maintaining physiological homeostasis, immune defense, and signal transduction. However, the complexity of plasma samples makes comprehensive protein analysis and high-precision identification particularly challenging, especially for low-abundance proteins, which are often masked by high-abundance ones and thus difficult to detect and analyze effectively.
One of the primary challenges in plasma proteomics is the efficient processing and extraction of proteins from samples. Conventional protein extraction and enzymatic digestion methods are often inadequate for plasma samples due to the overwhelming presence of high-abundance proteins (such as albumin), which dominate mass spectrometry signals and reduce the sensitivity of detecting low-abundance targets. To effectively address these challenges, researchers are in urgent need of a highly efficient, user-friendly enzymatic digestion method that can accurately target and process low-abundance proteins in plasma.
Product Overview
The plasma protein digestion kit by MtoZ Biolabs is a high-efficiency enzymatic digestion kit specifically optimized for plasma samples, designed for the rapid preparation and precise digestion of complex plasma proteins. Addressing common challenges such as the wide dynamic range of protein concentrations, interference from high-abundance proteins, and the presence of lipids and small molecular impurities, the kit integrates magnetic nanomaterials with an optimized enzymatic system to significantly enhance peptide yield and mass spectrometry compatibility.
The plasma protein digestion kit utilizes an improved digestion buffer along with high-purity trypsin to ensure effective breakdown of high-abundance proteins while increasing the detection probability of low-abundance proteins. The streamlined workflow and strong compatibility with multiple mass spectrometry platforms (including LC-MS/MS and MALDI-MS) make it an ideal choice for plasma proteomics research, preclinical biomarker discovery, and drug development sample preparation.
Product Notes
1. Lysis Buffer 1 must be freshly prepared before use by mixing components A and B.
2. Once sample preparation is complete, use the samples immediately or aliquot and store at -80°C.
3. The plasma protein digestion kit is designed to process 8 samples.
4. Avoid generating foam during operation, as it may affect protein binding efficiency.
5. It is recommended to use low protein-binding centrifuge tubes to collect the eluate, in order to prevent low-abundance proteins from adhering to tube walls and causing sample loss.
6. Protein concentrations can vary between plasma sample batches; therefore, it is advisable to perform a pilot experiment to determine the optimal dilution ratio and incubation time.
Product Details
|
Product Details |
Size |
Storage Conditions |
|
Wash buffer 1 |
2 mL×1 |
RT |
|
Wash buffer 2 |
8 mL×1 |
RT |
|
Lysis buffer 2 |
1.2 mL×1 |
RT |
|
Elution buffer 1 |
100 μL×1 |
RT |
|
Elution buffer 2 |
1 mL×1 |
RT |
|
Lysis buffer A |
3.75 mg×1 |
RT |
|
Lysis buffer B |
1 mL×1 |
RT |
|
Beads |
0.8 mg×1 |
4℃ |
|
Enzymes |
4 μg×1 |
-20℃ |
Protocol
The plasma protein digestion kit from MtoZ Biolabs adopts a modular design that integrates key steps including low-abundance protein enrichment, washing, lysis, and enzymatic digestion. The workflow is streamlined and time-efficient, making it suitable for routine or high-throughput mass spectrometry sample preparation.
1. Sample Preparation
Pipette 100-200 μL of plasma into a clean EP tube (dilution may be applied to optimize precipitation).
2. High-Abundance Protein Removal
(1) Add an appropriate volume of pre-washed magnetic beads to the plasma sample.
(2) Mix thoroughly and incubate on a rotator for 10-15 minutes.
(3) Use a magnetic rack to collect the beads and perform washing steps.
(4) Elute low-abundance proteins from the magnetic beads.
3. Low-Abundance Protein Digestion
(1) Dilute the protein solution using the digestion buffer and perform overnight enzymatic digestion.
(2) After digestion, collect the peptide solution using the elution buffer.
4. Digestion Termination and Sample Cleanup (Optional)
If necessary, terminate the reaction or proceed with desalting and other cleanup procedures according to experimental needs. The resulting peptides are ready for LC-MS/MS or other mass spectrometry analyses.
Features and Benefits
1. Optimized for Plasma Samples
The plasma protein digestion kit is specifically designed to address the complex background of plasma samples. It utilizes a selective precipitation strategy combined with mild enzymatic digestion conditions to effectively reduce interference from high-abundance proteins such as albumin, while ensuring complete release and efficient digestion of low-abundance proteins. This significantly enhances the depth of protein identification.
2. High-Efficiency Digestion System
Using high-activity, low-autolysis, sequencing-grade trypsin, coupled with precise pH and temperature control. This optimized system improves peptide generation quality, minimizes over-digestion and non-specific cleavage, and enhances the stability and quantitative accuracy of downstream mass spectrometry analysis.
3. Excellent Mass Spectrometry Compatibility
The resulting peptides are highly compatible with mainstream platforms such as LC-MS/MS (DDA/DIA modes), PRM, and MALDI-TOF. The plasma protein digestion kit supports qualitative identification, quantitative analysis, and post-translational modification studies, particularly in the discovery of biomarkers in plasma proteomics.
FAQs
Q1: What Types of Plasma Samples Is the Kit Suitable for?
A1: The plasma protein digestion kit is suitable for various types of plasma samples derived from healthy individuals, disease models, or preclinical studies. Whether you're targeting high-abundance proteins such as albumin or low-abundance proteins like cytokines and hormones, the kit delivers efficient protein digestion to support comprehensive proteomic analysis. Its high compatibility allows it to handle complex plasma matrices, providing broad proteome coverage across diverse sample types.
Q2: Which Mass Spectrometry Platforms Is the Kit Compatible with?
A2: The plasma protein digestion kit is highly compatible with multiple mass spectrometry platforms, including LC-MS/MS (DDA/DIA modes), MALDI-MS, and PRM. Whether you are performing qualitative identification, quantitative profiling, or post-translational modification studies, the kit ensures stable and high-quality sample preparation, delivering reliable accuracy and sensitivity for downstream MS analysis.
