Fungus (Flesh-Like Fungi) Protein Extraction Kit

Product Name	Fungal (Flesh-Like Fungi) Protein Extraction Kit	Storage Conditions	4℃
Kit Components	Extraction Buffer 1, Extraction Buffer 2, PMSF	Applicable Organisms	Unicellular fungal samples
Product Type	Kit	Shelf Life	1 year
Product Size	48T/box	Compatible Applications	Protein identification, proteomics, IP, WB

  

Protein is one of the most essential functional molecules in living organisms, extensively involved in cellular metabolism, signal transduction, immune responses, and biosynthesis. Studying protein expression and function not only helps scientists deepen their understanding of biological mechanisms but also drives technological advancements across various fields such as medicine, food science, biomaterials, and environmental science. Flesh-like fungi, also known as macrofungi, refer to fungal groups that possess visible mycelia or fruiting bodies, including fresh shiitake mushrooms, Ganoderma lucidum (reishi), Cordyceps sinensis, matsutake, Tremella, and others. These fungi are of significant value in biomedical research, medicinal active compound extraction, the food industry, and ecological system balance.

 

Due to the robust cell wall structure of flesh-like fungi, which is rich in chitin and polysaccharides, conventional protein extraction methods often fail to efficiently lyse the cells, resulting in low protein yield and reduced protein activity. The fungus (flesh-like fungi) protein extraction kit uses an optimized chemical lysis system tailored to the cellular characteristics of macrofungi. It enables efficient and gentle disruption of cell walls and releases high-quality intracellular proteins, providing researchers with a reliable and effective solution for protein extraction.

 

Product Overview

The fungus (flesh-like fungi) protein extraction kit from MtoZ Biolabs is specifically designed for macrofungi such as mushrooms, Ganoderma, Cordyceps, Tremella, and matsutake. It employs a mild yet efficient lysis method that allows for rapid extraction of intracellular proteins. The kit is suitable for protein extraction from flesh-like fungi and supports various downstream applications including Western blotting, SDS-PAGE, ELISA, and LC-MS/MS-based proteomics. It is widely applicable in proteomics, immunology, biopharmaceutical research, and food science.

 

Protocol

The fungus (flesh-like fungi) protein extraction kit has been optimized to utilize a simplified chemical lysis process, enabling efficient and straightforward extraction of various flesh-like fungi. It is suitable for different laboratory environments and experimental scales. Below is the recommended standard procedure, which can be adjusted based on actual experimental needs.

1. Fungal Sample Preparation

(1) Grind an appropriate amount of fresh fungal tissue into powder using liquid nitrogen.

(2) Add Protein Extraction Buffer 1 (provided in the kit) at the recommended ratio and thoroughly resuspend the fungal powder.

Note: It is recommended to add 1 mL of extraction buffer per 200 mg of fungal tissue.

(3) Centrifuge at 15,000 ×g at low temperature for 10 minutes and collect the protein-containing supernatant.

(4) Add 4 volumes of Extraction Buffer 2 (provided in the kit) and vortex thoroughly for 10 minutes.

(5) Add 5 volumes of pre-chilled Extraction Buffer 3 (provided in the kit) to precipitate the proteins.

(6) Incubate the mixture at -20°C for 16 hours.

(7) Centrifuge again at 15,000 ×g at 4°C for 10 minutes.

(8) Wash the pellet 2-3 times with pre-chilled Extraction Buffer 3.

(9) Further wash the pellet twice using pre-chilled 70% ethanol and 80% acetone.

(10) Air-dry the pellet at room temperature and store the dried protein at -80°C.

  

2. Protein Quantification and Storage

(1) Use an appropriate protein quantification method to determine protein concentration and ensure it meets the experimental requirements.

(2) For short-term storage (1-2 weeks), store the extracted protein at 4°C.

(3) For long-term storage (>2 weeks), it is recommended to store the samples at -80°C to prevent protein degradation.

 

Figure 1. Extraction Effect Diagram of Fungus (Flesh-Like Fungi) Protein Extraction Kit.

 

3. Experimental Applications

(1) The extracted proteins can be directly used for SDS-PAGE, Western blot, ELISA, LC-MS/MS, enzyme activity assays, and other downstream experiments.

The entire procedure requires no additional advanced equipment, significantly improves experimental efficiency, and minimizes sample loss.

 

Product Notes

1. Avoid prolonged incubation at high temperatures to prevent protein degradation.

2. To reduce the risk of protein degradation, avoid repeated freeze-thaw cycles; aliquot samples for storage when possible.

3. Optimize the volume of lysis buffer according to experimental needs to improve protein extraction efficiency.

 

Features and Benefits

1. Efficient and Gentle Lysis

Utilizes an optimized chemical lysis system to avoid mechanical damage to proteins, enabling rapid and efficient disruption of the cell walls of flesh-like fungi, thereby releasing high-quality intracellular proteins while minimizing protein denaturation and degradation.

 

2. High Reproducibility and Data Consistency

The standardized and optimized lysis system ensures consistent and stable results across different experimental batches.

 

3. Compatible with Various Downstream Applications

The high-yield protein extracts are suitable for SDS-PAGE, Western Blot, ELISA, LC-MS/MS, enzyme activity assays, and other experiments to meet diverse research needs.

 

4. Simple Operation

Ready-to-use reagents eliminate the need for complex instruments, offering a straightforward workflow with minimal sample loss.

 

Applications

1. Proteomics Research

(1) The fungus (flesh-Like fungi) protein extraction kit is suitable for LC-MS/MS analysis to profile protein expression and identify differentially expressed proteins in flesh-like fungi, supporting comprehensive proteomics research.

 

2. Food Science and Functional Food Development

(1) Applicable to the analysis of edible fungal proteins, aiding studies on functional proteins from fungi such as matsutake, tremella, and mushrooms, and optimizing food processing methods.

(2) Enables research on specific proteins in flesh-like fungi, such as nutritional proteins and polysaccharide-binding proteins, contributing to the development of functional foods.

 

3. Medicinal Bioactivity Research

(1) Facilitates the study of functional proteins from medicinal fungi such as Ganoderma lucidum and Cordyceps sinensis, allowing screening of bioactive proteins and research on antifungal drugs to accelerate the application and development of medicinal fungi.