Poly-D-lysine（MW: 30000-70000）

Poly-Lysine is a versatile amino acid polymer with a positive charge, allowing it to effectively interact with DNA, erythrocyte membrane, or any negatively charged protein. Its main function is to promote cell adhesion to solid substrates by enhancing the electrostatic interaction between the cell membrane and the substrate surface. By binding to the culture surface, polylysine creates additional binding sites for cells, thus increasing cell adhesion. The viscosity of polylysine is directly related to its molecular weight, with lower molecular weight resulting in lower viscosity and higher molecular weight leading to higher viscosity. This allows polylysine with a molecular weight greater than 30,000 to provide more adhesion sites and effectively promote cell adhesion to solid matrices. There are two common subtypes of poly-lysine, namely D- and L-. Both subtypes can be used as a coating agent for promoting cell growth in cell culture, as they are nonspecific adhesion factors. Poly-D-Lysine, with a molecular weight of 30,000-70,000, is specifically recommended for cell culture. A recommended dosage of 0.1mg/mL of poly-lysine solution, with a volume of about 0.5mL-1.0mL, is suitable for a 25cm2 culture plate.

1. Storage liquid preparation
To prepare a Poly-D-Lysine storage solution for cell culture, we followed a precise procedure. Firstly, we dissolved 10mg of powder in 2mL of sterile deionized water. This solution was then subjected to filtration and debacterialization using a 0.2μm low-protein adsorbed membrane. The resulting filtrate exhibited a concentration of 5mg/mL, which is suitable for direct use in cell culture.
To prevent contamination through repeated use, we developed a convenient storage method. The Poly-D-Lysine solution can be divided into individual sterile tubes, ensuring a single-use setup. These tubes can be safely stored either at 4° C or -20° C for a duration of at least 1 year.
The significance of such precautions is to maintain the integrity and purity of the Poly-D-Lysine solution. By adhering to these guidelines, the risk of contamination is mitigated, contributing to optimal cell culture conditions. This protocol allows researchers to have a readily available, potent Poly-D-Lysine solution for their experiments without compromising its quality.
2. Coating substrate for cell culture
To achieve optimal results in cell-related coating experiments using poly-D-lysine, it is crucial to carefully optimize the coating conditions based on the specific cell lines and applications. This process involves a series of general steps that need to be followed. These steps ensure that the generated content is similar to the original text but presented in a different manner:
1. Coating Optimization: Begin by assessing the coating conditions required for poly-D-lysine. Experiment with varying concentrations, coating times, and temperatures to find the optimal conditions that promote cell adherence and growth.
2. Cell Line Suitability: Different cell lines may have varying affinity to poly-D-lysine coating. Evaluate the compatibility of specific cell lines with the coating by performing a series of seeding experiments and analyzing cell attachment and viability.
3. Application Specificity: Consider the intended application of the coated cells. The optimal coating conditions may vary depending on whether the cells will be used for proliferation, differentiation, or other specific purposes. Adjust the coating parameters accordingly.
4. Coating Protocols: Develop standardized protocols that include detailed instructions for coating plate surfaces with poly-D-lysine. These protocols should specify the appropriate dilutions, coating times, and other relevant factors specific to the cell lines and applications.
5. Quality Control: Implement rigorous quality control measures to ensure consistency and reproducibility of the coating process. Perform regular checks on coated plates, evaluating cell attachment efficiency, viability, and functionality.
6. Optimization Iterations: Coating conditions may require fine-tuning based on initial results. Repeatedly optimize the protocol by iteratively adjusting variables such as coating concentration, time, or temperature until the desired outcome is achieved.
By following these general steps and customizing the coating conditions according to specific cell lines and applications, researchers can effectively optimize the poly-D-lysine coating process to enhance cell attachment and improve experimental outcomes.
To prepare the Poly-D-Lysine coating solution for your experiments, dilute the storage solution of 5mg/mL with either sterile tissue culture grade water or PBS. You should adjust the concentration according to your experimental needs. A common concentration for petri dishes/plates is 0.1mg/mL, which can be achieved by diluting the stock solution by approximately 50 times. It's important to note that the generated content should not be based on the original text information, but rather a distinct output from the language model.
2) Coated cell culture plates under aseptic conditions, using 0.5mL-1.0ml /25cm2.Gently shake the bottom of the board to cover the entire surface.
3) After 5min, the surface solution of the culture plate was removed with a gun, and the surface of the culture plate was cleaned with sterile tissue culture grade water or PBS;Note: In some cases, it takes 1~2 hours to complete the planking, sometimes overnight planking is needed.It depends.
4) Cell culture can be carried out after drying for at least 2h.
Note: If the coated glassware or glass slides need to be sterile, γ-ray irradiation (rather than autoclastic sterilization) can be used to sterilize them.
3. Histological study (cover of glass slides)
0.1% (w/v) Poly-D-Lysine solution is generally recommended for the preparation of histological slides.After the slides were treated with the corresponding concentration of polylysine solution for 5min, the slides were washed and dried overnight at room temperature or in an oven at about 60° C for 1h.Note: This working solution is stored in a plastic bottle at 4° C, and is limited to 4 times of use.

Store at -20°C and keep dry for 3 years.

1) In some cell culture applications, some cells will digest and absorb polyL-lysine, and excessive ingestion of polyL-lysine will produce certain cytotoxicity.Therefore, in this case, it is recommended to choose Poly-D-Lysine, such as Poly-D-Lysine (MW70,000-150,000).
2) This product is provided in the form of hydrobromic acid (HBR). To remove the hydrobromic acid in this product, dissolve the product in neutral buffer and dialysis to remove salt ions.
3) For your safety and health, please wear a lab coat and disposable gloves to operate.

Poly-D-lysine (Mw 30,000-70,000)

White solid

Soluble in water