Other Tool Enzymes

This Recombinant Proteinase K is a genetically engineered recombinant serine protease. It is produced by introducing the functional gene of fungal protease K into the Pichia pastoris expression system, through high-density fermentation and multiple purification processes.

This enzyme has broad-spectrum protein hydrolysis ability and can efficiently cleave the carboxyl-terminal peptide bonds of most amino acids. It has a higher affinity for aliphatic and aromatic amino acid substrates. The product has undergone strict quality control screening and is free from DNase, RNase, endonucleases, and exonucleases, with a purity that meets the standards for high-end nucleic acid extraction, library construction sequencing, and molecular cloning experiments.

Compared to ordinary proteinase K, the main feature of this product is that it can be completely heat-inactivated. It can be completely deactivated by heating at 55℃ for 10 minutes. No additional enzyme purification is required, and it supports single-tube integrated sample processing, significantly simplifying the molecular experiment process.

1. Complete thermal inactivation, simple experimental process

The common proteinase K still retains residual activity after heating, which interferes with downstream experiments. This product has excellent thermal stability and can completely inactivate at low temperatures, achieving sample digestion - heating inactivation - direct single-tube operation, eliminating the need for magnetic bead purification and repeated elution steps, and preventing enzyme residue contamination.

2. Stable enzyme activity, broad substrate compatibility

After gene site-directed modification, while retaining the rapid thermal inactivation characteristic, it completely inherits the hydrolytic activity and substrate specificity of wild-type proteinase K, and can efficiently degrade various biological samples containing impurities and residual enzymes.

3. Ultra-pure without contaminating enzymes, zero interference in experiments

The strict protein purification and quality control system has no residual impurities of various nucleases and proteases. The experimental background is clean and stable, suitable for high-precision experimental scenarios such as sequencing, PCR amplification, and micro-sample preparation.

4. Strong adaptability, extremely high system compatibility

Tolerant to complex reaction environments, it can work stably in different salt concentrations, conventional buffers, EDTA, DTT, and detergent systems; the optimal activity range is 20–40℃, pH 7.0–9.5, and it can still maintain efficient hydrolysis ability in SDS and low-concentration chelating agent environments.

5. Suitable for micro and high-throughput experiments

The entire process is single-tube operation, minimizing the loss of nucleic acid samples during transfer, significantly improving the recovery rate of low starting volume samples, and suitable for automated extraction equipment and high-throughput sample screening systems.

Notes for Experimental Use

1. Optimal reaction conditions: pH 7.0–9.5, 20–40°C. Long-term exposure above 35°C will reduce enzyme activity stability;

2. Tolerates chelator systems containing ≤10 mM EDTA, suitable for conventional nucleic acid extraction;

3. Urea concentration >2 M in the system inhibits enzyme activity; high urea environments should be avoided;

4. Serine protease inhibitors such as PMSF block the enzymatic reaction. Do not add them to the experimental system;

5. Low-concentration detergents can enhance performance: ≤1% SDS improves hydrolysis efficiency; SDS within 2.5% and Triton X-100 within 1.5% do not inhibit enzyme activity.

Buffer system: 20 mM Tris-HCl, 1 mM CaCl₂, 50% Glycerol, pH 7.4 (25°C)

1. Long-term preservation: It is recommended to store at -25℃ to -15℃ in a sealed freezer for long-term stable preservation;

2. Short-term preservation: For routine use within one year, it can be stored at 4℃;

3. Preservation suggestions: Do not repeatedly freeze and thaw. After receiving the product, aliquot and store as needed to maximize the retention of activity; the freeze-dried powder form can be stored for a long time under dry and sealed conditions at 4℃.

1. Nucleic acid sample preparation: Extraction of genomic DNA and total RNA, digestion of sample to remove non-specific proteins, thorough removal of endogenous RNase and DNase interference;

2. PCR product optimization: Degradation of residual polymerase proteins during amplification, elimination of enzyme residue influence, and improvement of molecular cloning success rate;

3. Pathological in situ experiment: Pre-treatment and digestion of tissue sections, adaptation of in situ hybridization experimental system;

4. High-throughput library construction sequencing: Removal of protein impurities during DNA/RNA library construction to ensure library quality;

5. Biological pharmaceutical detection: Removal of non-specific protein impurities in biological samples and preparations to meet pharmaceutical quality inspection standards;

6. Microscale / Automated experiments: Adaptation to low-concentration nucleic acid samples and high-throughput automated extraction platforms.