The MtoZ Biolabs' microtissue protein digestion kit is specifically designed for tissue samples with low starting material and complex backgrounds. It optimizes digestion efficiency and peptide recovery, making it compatible with downstream LC-MS/MS workflows. The kit is particularly suitable for proteomic analysis of tissue sections, micro-needle biopsies, and other small-volume tissue samples, significantly improving the depth and reproducibility of downstream protein identification. It is widely applicable in basic medical research, oncology studies, and clinical translational research.
As proteomics expands toward precision clinical samples and spatially resolved research, researchers increasingly face challenges in protein extraction and digestion from micro-tissue samples. Examples include FFPE tissue sections, laser capture microdissection (LCM) samples, needle biopsy tissues, and tissues from small animal models. In these sample types, low protein content, complex background, and unstable digestion efficiency are common issues. These challenges directly impact the number of proteins identified, quantification accuracy, and inter-batch reproducibility in mass spectrometry data.
Traditional digestion methods (such as FASP and in-solution digestion) perform adequately with conventional samples but often encounter issues when applied to micro-tissue samples, including low protein recovery, incomplete digestion, and poor reproducibility. In addition, residual interfering substances in the samples (such as lipids, pigments, nucleic acids, and salts) frequently affect subsequent MS signal quality.
Product Overview
MtoZ Biolabs' microtissue protein digestion kit provides a fully optimized workflow from tissue lysis to enzymatic digestion, with features of efficient digestion, low sample loss, and compatibility with NanoLC-MS/MS platforms, making it suitable for studies in tissue proteomics, spatial proteomics, and low-abundance protein analysis. The kit is compatible with tissues from multiple species, including human, mouse, and zebrafish, and meets the urgent needs of researchers for high-quality processing of micro-scale samples, which is especially suitable for highly complex micro-tissue samples that are difficult to handle using conventional methods, and can be used as a standardized solution for converting tissue into peptides.
Product Details
|
Product Details |
Size |
Storage Conditions |
|
Lysis buffer |
10 mL×1 |
-20℃ |
|
Beads |
200 μL×1 |
4℃ |
Protocol
The microtissue protein digestion kit from MtoZ Biolabs has undergone workflow integration and parameter optimization, enabling easy completion of all steps from tissue lysis to peptide purification. It is particularly suitable for samples with limited starting material and complex backgrounds. The recommended protocol is as follows:
1. Tissue Lysis
(1) Weigh 1-10 mg of wet tissue sample and add an appropriate volume of lysis buffer. Homogenize thoroughly.
2. Alkylation
(1) Add the corresponding volume of alkylation reagent to block free thiol groups.
3. Magnetic Bead Binding and Digestion
(1) Add pre-treated magnetic beads to bind target proteins.
(2) Wash to remove impurities.
(3) Add trypsin and incubate at 37°C for 14-16 hours.
4. Peptide Recovery and Purification
(1) After digestion, use magnetic separation to collect the supernatant, which contains the peptide solution.
(2) Perform StageTip purification and desalting.
Features and Benefits
- Integrated processing workflow: tissue lysis + alkylation + magnetic bead-based digestion
- Strong compatibility with micro-scale samples: stable processing of 1-10 mg wet tissue without protein quantification or sample transfer loss
- Mass spectrometry-compatible buffer system: digestion products are directly applicable to NanoLC-MS/MS analysis, compatible with mainstream platforms such as Orbitrap, TripleTOF, and timsTOF
- Broad tissue type compatibility: suitable for muscle, liver, brain tissue, tumor tissue, and FFPE samples
- High inter-batch stability: ideal for large-scale proteomics study designs (e.g., TMT/iTRAQ)
Applications
The following research areas can greatly benefit from the microtissue protein digestion kit's high digestion efficiency and low sample loss characteristics:
1. Clinical Minimally Invasive Sample Studies
Proteomics sample preparation for small-volume clinical samples such as fine-needle aspiration and core biopsies.
2. Proteomic Analysis of Small Animal Model Tissues
Suitable for microdissected tissues from brain, liver, kidney, etc., in models such as mice, zebrafish, and monkeys.
3. Assisted Digestion for Spatial Proteomics
Quantitative proteomic analysis of tissue regions after laser capture microdissection (LCM).
4. Low-Abundance Protein Screening and Biomarker Discovery
Enhances recovery and detection rates of low-concentration proteins, supporting research on early-stage disease biomarkers.
FAQs
Q1: What Is the Minimum Tissue Amount Required for the Kit?
A1: The recommended starting amount is 1 mg of fresh or frozen tissue. However, under optimized conditions, the microtissue protein digestion kit can also be used for samples of less than 1 mg. MtoZ Biolabs has developed a closed reaction system specifically optimized for microtissue samples, ensuring efficient protein recovery even at ultra-low input levels.
Q2: Is the Microtissue Protein Digestion Kit Suitable for Tissue Samples with High Fat Content?
A2: Yes. The kit contains an optimized lysis buffer with effective defatting capacity. For high-fat content samples such as brain or adrenal gland, it is recommended to combine with additional preprocessing steps to further enhance lysis efficiency.
