Fruit, as a critical reproductive organ of plants, plays not only a major role in species propagation but also serves as a central hub in plant growth, metabolism, and environmental adaptation. Fruits are rich in various biological macromolecules, including proteins, enzymes, lipids, and carbohydrates, all of which are essential for vital activities such as nutrient storage and seed development. With ongoing advances in plant biology, functional genomics, and food science, the functional roles and dynamic changes of fruit proteins have increasingly become research hotspots, particularly regarding disease resistance, stress tolerance, and the ripening process. As such, extracting high-quality proteins from fruit and conducting in-depth analyses has become an indispensable step in fruit-based plant research. However, the extraction of high-quality proteins from plant fruits still faces many challenges. For example, fruits contain a variety of polyphenols, esters, and polysaccharide compounds, due to the strict separation conditions, the activity of endogenous enzymes is easy to destroy the stability of fruit protein, the isoelectric point range of protein is wide, and aggregation precipitation is easy to occur under certain conditions.
To address these challenges, MtoZ Biolabs has specifically developed the plant fruit protein extraction kit which designed to provide a simple, efficient, and reliable solution for extracting proteins from plant fruits. By carefully optimizing the lysis buffer, protease inhibitor system, and detergent components, the kit effectively overcomes various difficulties encountered during fruit protein extraction. It is broadly applicable to a wide range of fruit types, enabling researchers to obtain high-purity, high-activity protein samples in a short amount of time, and provides solid technical support for downstream research.
Product Overview
The plant fruit protein extraction kit is a highly efficient kit specifically developed to address the challenges of protein extraction from plant fruits. It features a lysis buffer formula optimized for fruit tissues, enabling rapid disruption of fruit cell walls and membranes under mild conditions to release intracellular proteins. The kit includes protease inhibitors that effectively suppress endogenous protease activity in fruits, preventing degradation of target proteins during extraction. Additionally, the unique detergent components efficiently remove interfering substances such as pectin, sugars, and lipids, ensuring the extracted proteins are of high purity and stability.
Plant fruit protein extraction kit is suitable for a variety of plant fruits, including tomato, citrus, grape, apple, and peach, and enables rapid protein extraction under standard laboratory conditions. The resulting protein samples are compatible with a wide range of downstream applications, including Western blotting, ELISA, mass spectrometry (LC-MS/MS), immunoprecipitation, and enzyme activity assays, meeting the needs of research across botany, nutrition science, and food science.
Product Details
|
Product Details |
Size |
Storage Conditions |
|
Plant Fruit Lysis Buffer |
40 mL |
4℃ |
|
Protease Inhibitor (100×) |
0.5 mL |
-20℃ |
Protocol
Below is the standard operating procedure for the plant fruit protein extraction kit. Users may adjust the steps as needed based on the fruit type and specific experimental requirements:
1. Sample Preprocessing
Select fresh, healthy plant fruits and remove surface debris. Cut the fruit into small pieces to facilitate grinding and lysis. For fruits with tough skins, cryogenic grinding with liquid nitrogen is recommended.
2. Lysis Buffer Preparation
a. Thaw the plant fruit lysis buffer and place it on ice. Add protease inhibitor (100×) and mix well (add 10 μL of protease inhibitor per 1 mL of lysis buffer).
b. Add 3-4 volumes of lysis buffer relative to the fruit tissue volume. Incubate on ice or at 4°C for 30-60 minutes with shaking, or for 3-4 hours without shaking.
c. Mechanically disrupt the sample using a homogenizer or ultrasonic disruptor.
3. Centrifugation and Supernatant Collection
a. Centrifuge at 10,000-12,000 ×g for 10-15 minutes at 4°C to pellet cell debris and fibrous materials.
b. Carefully transfer the supernatant to pre-chilled tubes. This is the extracted fruit protein solution.
c. Aliquot and store at -80°C. For higher purity, additional centrifugation or membrane filtration can be performed.
4. Protein Precipitation (Optional)
Add pre-chilled acetone and incubate at -20°C for at least 1 hour. Centrifuge at 12,000 ×g for 15 minutes, air-dry the pellet, and reconstitute in an appropriate buffer.
5. Protein Quantification and Downstream Applications
Quantify the extracted protein using a standard method such as BCA. The protein samples are suitable for various downstream analyses, including Western blotting, ELISA, LC-MS/MS, and enzymatic assays.
This workflow ensures efficient extraction of fruit proteins and is straightforward to perform without requiring specialized instruments, making it well-suited for standard laboratory environments.
Features and Benefits
1. High-Efficiency Lysis
The lysis is specially optimized to effectively disrupt fruit cell walls under mild conditions and release intracellular proteins without causing excessive damage. This results in high extraction efficiency.
2. Protease Inhibition System
A built-in multi-component protease inhibitor system effectively suppresses endogenous protease activity in fruit tissue, preventing protein degradation and ensuring the integrity and activity of extracted proteins.
3. Optimized Detergent Composition
The unique detergent formulation efficiently removes excess sugars, lipids, pectins, and other interfering substances from fruit tissues, minimizing impurities and ensuring the extraction of high-purity proteins.
4. Streamlined Protocol
The plant fruit protein extraction kit offers ready-to-use reagents with a simplified workflow, allowing for rapid protein extraction in a short time, minimizing handling steps and experimental error, and saving valuable lab time.
5. High Compatibility
The extracted fruit proteins are broadly compatible with various downstream applications, including Western blotting, ELISA, mass spectrometry (LC-MS/MS), and enzyme activity assays, making it suitable for multidisciplinary research.
6. Applicable to a Wide Range of Fruit Types
The plant fruit protein extraction kit is compatible with various plant fruits, such as tomato, citrus, grape, apple, and peach, demonstrating strong adaptability for different fruit sample extraction needs.
Applications
1. Plant Growth and Development Studies
Analyze protein expression changes in fruits at various developmental stages to uncover the molecular mechanisms underlying plant growth, fruit development, and ripening.
2. Environmental Adaptation and Stress Resistance Research
Monitor changes in antioxidant enzymes and stress-responsive proteins in fruits under environmental stress conditions (e.g., drought, salinity, pests), revealing plant adaptation strategies in adverse environments.
3. Food Nutrition and Quality Assessment
Investigate nutrient components and enzyme activities in fruits to evaluate their stability and transformation under different storage and processing conditions, supporting food quality control and processing optimization.
4. Gene Function and Transgenic Plant Research
Conduct comparative proteomic analysis of fruits from different genetic backgrounds (e.g., transgenic plants) to identify functional proteins associated with traits, disease resistance, and nutritional value, providing reference data for plant breeding.
5. Functional Analysis of Plant Proteins
Study key enzymes and transport proteins in fruit tissues to elucidate their roles in metabolism, signal transduction, and fruit maturation, shedding light on their essential functions in plant life processes.
FAQs
Q1: How Can Protein Degradation Be Avoided during Extraction?
A1: Protein degradation primarily results from endogenous protease activity and can be prevented by: 1. Low-temperature operation: Perform the entire process on ice and pre-chill all reagents and instruments. 2. Protease inhibitors: Add broad-spectrum inhibitors to suppress protease activity. 3. Rapid processing: For fresh samples, complete extraction within 30 minutes of collection. For frozen samples, avoid repeated freeze-thaw cycles.
Q2: What Might Cause Low Protein Concentration in the Extract, and How Can it Be Resolved?
A2: 1. Insufficient sample volume: Use larger quantities for low-protein tissues like fruit; extend grinding time for fibrous samples. 2. Incomplete lysis: Use intermittent homogenization or supplement with ultrasonic disruption to release tightly bound proteins. 3. Incorrect reagent ratio: For high-starch samples (e.g., potatoes), reduce the volume of lysis buffer to concentrate the protein.
Q3: Can the Extracted Protein Be Directly Used for Enzymatic Activity Assays or Western Blotting?
A3: 1. Enzyme activity assays: Yes, the supernatant can be used directly, but ensure the lysis buffer is free of denaturants (e.g., SDS). 2. Western blotting: If the sample contains high salt or pigments, it is recommended to purify using acetone precipitation or dialysis. 3. Mass spectrometry: Quantify using the BCA method and confirm purity; if necessary, use desalting columns to remove interfering small molecules.
