In the human immune system, MHC-I (major histocompatibility complex class I) molecules play a critical role in antigen presentation. They present endogenous peptides (such as those derived from viral infections or tumor cells) to CD8+ T cells, serving as a key component in immune surveillance and response. These peptides presented by MHC-I molecules are referred to as MHC-I peptides. To further understand peptide-receptor interactions, it is necessary to enrich MHC-I peptides from the cell surface or cell lysates. This work is essential for antigen epitope discovery, vaccine development, and tumor immunotherapy.
However, the enrichment of MHC-I peptides faces several challenges, including low peptide abundance, high background interference, and instability of extraction methods. Traditional enrichment techniques often rely on complex experimental procedures and lengthy operations. To improve enrichment efficiency, reduce background noise, and simplify the workflow, MtoZ Biolabs has developed a human MHC class I peptide enrichment kit to provide researchers with a convenient and efficient solution for enriching MHC-I peptides.
Product Overview
The human MHC class I peptide enrichment kit from MtoZ Biolabs is specifically designed for the efficient enrichment of human MHC-I peptides. It employs a highly specific antibody-beads binding technique, enabling rapid and accurate enrichment of MHC-I peptides from complex samples. The core advantages of the kit lie in its ability to significantly reduce background noise, improve peptide recovery rate, and shorten the experimental cycle. It is helpful for researchers to quickly extract MHC-I peptides from cell or tissue lysates, providing high-quality samples for subsequent mass spectrometry analysis, epitope discovery, and immune response studies.
Product Details
|
Product Details |
Size |
Storage Conditions |
|
Lysis buffer |
10 mL×1 |
4℃ |
|
Binding buffer |
5 mL×1 |
4℃ |
|
Elution buffer |
20 mL×1 |
4℃ |
|
Wash buffer A |
20 mL×1 |
4℃ |
|
Wash buffer B |
20 mL×1 |
4℃ |
|
Wash buffer C |
20 mL×1 |
4℃ |
|
Antibody |
50 µL×1 |
-20℃ |
|
Beads |
2 mL×1 |
4℃ |
|
Column tubes |
4 pcs |
RT |
Protocol
The human MHC class I peptide enrichment kit from MtoZ Biolabs adopts standardized experimental procedures, enabling researchers to quickly get started and obtain high-quality enrichment results. The following is an overview of the recommended workflow:
1. Cell Lysis and Sample Preparation
Add Lysis Buffer to the cell or tissue samples and perform pretreatment according to the protocol to extract proteins.
Note: It is recommended to verify the presence of MHC-I peptides in the extracted proteins using the Western blot (WB) method.
2. Protein Purification
Add beads to a column tube and wash three times, then add the protein solution for purification.
3. Immunoaffinity Enrichment
(1) Transfer beads to a new tube, wash, and add an appropriate amount of antibody. Incubate at 4°C with rotation for 4 hours.
(2) Wash the antibody-conjugated beads multiple times with wash buffer to remove nonspecifically bound impurities.
(3) Add Elution Buffer and perform four elution steps.
Note: It is recommended to verify the enriched immunopeptides after elution using the WB method to ensure enrichment efficiency.
4. Desalting
Remove salts from the eluted solution to keep the peptides pure.
5. Downstream Analysis
The extracted peptides can be analyzed by mass spectrometry to further identify peptide sequences, or used for immune response assays and related applications.
Features and Benefits
The human MHC class I peptide enrichment kit from MtoZ Biolabs offers multiple advantages:
1. High-Efficiency Enrichment
The combination of specific antibodies with beads significantly improves enrichment efficiency. It enables efficient extraction from low-abundance peptides while minimizing background interference to the greatest extent.
2. Simplified Workflow
Standardized operating procedures eliminate complex manual steps, allowing researchers to complete high-quality enrichment in a short period of time.
3. Strong Compatibility
Applicable to various sample types, including cells and tissues. Human MHC class I peptide enrichment kit is suitable for both basic research and translational clinical studies.
4. Optimized Reagent Formulation
Each component in the kit is optimized to ensure high stability and sensitivity during the enrichment process, reducing interference from background noise.
Applications
The human MHC class I peptide enrichment kit is widely applicable across various fields, particularly in the following research areas:
1. Tumor Immunology Research
In cancer immunotherapy, MHC-I peptides play a vital role in tumor immune surveillance and immune evasion. By enriching MHC-I peptides, researchers may identify potential cancer immunoepitopes for vaccine development and immunotherapy evaluation.
2. Antigen Presentation Analysis
Used to detect and analyze antigenic peptides presented by MHC-I molecules, assisting researchers in identifying critical immune epitopes, thereby advancing vaccine development and infection research.
3. Immune Evasion Mechanism Studies
In the study of autoimmune diseases and immune evasion mechanisms, analyzing changes in MHC-I peptides can reveal immune system responses under pathological conditions.
4. Biomarker Discovery for Immunotherapy
Enrichment and analysis of MHC-I peptides can aid in identifying biomarkers for immunotherapy, enhancing the specificity and efficacy of treatment strategies.
FAQs
Q1: Can the Human MHC Class I Peptide Enrichment Kit Be Used for Low-Abundance Samples?
A1: Yes. The kit is optimized for high-efficiency enrichment of MHC-I peptides from low-abundance samples, making it suitable for most biological materials.
Q2: How Is Enrichment Accuracy Ensured?
A2: The kit utilizes high-affinity antibodies conjugated to beads and incorporates multiple washing steps to effectively remove nonspecific binders, thereby ensuring enrichment specificity and accuracy.
Q3: What Types of Samples Are Compatible with the Kit?
A3: The human MHC class I peptide enrichment kit is compatible with a variety of sample types, including cultured cells and tumor tissues.
Q4: Is the Enriched Peptide Material Compatible with Mass Spectrometry Platforms?
A4: Yes. The enriched MHC-I peptides can be directly analyzed by mass spectrometry for sequence identification and downstream applications.
Q5: How Long Can the Enriched Peptides Be Stored?
A5: It is recommended to store enriched peptides at -20°C. Under proper conditions, they can be stably preserved for several weeks.
