Cell Electrofusion Buffer

Our product is specifically optimized for hybridoma technology, with the core advantages of "High Efficiency, Stability, and Compatibility." The precise low-conductivity formulation minimizes Joule heating during electrofusion, ensuring high cell viability and thereby achieving higher fusion efficiency. Furthermore, our stringent process control guarantees high batch-to-batch consistency, leading to strong reproducibility of experimental results. More importantly, this product has been tested and is compatible with mainstream electroporators on the market, saving you the hassle of adaptation and optimization.

No need for complex parameter adjustments. The buffer provides a standard conductivity and osmolarity environment, creating ideal and stable conditions for electrofusion. You can directly use the standard electrofusion programs "Splenocyte/SP2/0" or "Splenocyte/P3" recommended in our protocol on your instrument. This simplifies the workflow and lowers the threshold to use.

Thorough cleaning and pre-treatment of the fusion chamber are crucial for success, focusing on removing ionic contamination and maintaining a low temperature. Key steps include:

1. Pre-cooling: Place the fusion chamber in a 4°C refrigerator or on ice for at least 30 minutes in advance.

2. Cleaning and Disinfection: Soak it in 75% ethanol for 10 minutes, then rinse twice with sterilized deionized water.

3. Equilibration: Finally, it is essential to rinse the chamber twice with this buffer (Cell Electrofusion Buffer) to ensure that the conductivity of the fusion chamber environment is consistent with that of the cell suspension.

We strongly recommend completing the electrofusion operation within 1 hour, and ideally within 10 minutes, after cell washing. There are two main reasons:

1. Maintaining Cell Viability: Although this buffer is non-toxic, it does not contain any nutrients. Cells suspended in it for too long (more than 1 hour) can lead to decreased cell viability.

2. Ensuring Fusion Efficiency: If cells stay in the buffer for too long (especially more than 30 minutes), their electrophysical properties may change, significantly reducing fusion efficiency. Additionally, the fusion program should be initiated within 30 seconds after adding the cells to the chamber to prevent sedimentation from affecting the effect.

We recommend using a 1:1 ratio of splenocytes to SP2/0 cells for fusion to achieve optimal hybridoma formation.

After the electric fusion program is completed, please follow these steps:

1. Allow to stand: Allow the fused cells to stand in the fusion chamber for 2-3 minutes for a stable recovery process for the fused cell membranes.

2. Transfer and Incubation: Gently collect the cells into a pre-prepared fully fused culture medium.

3. Recovery Culture: Incubate in a 37°C, 5% CO₂ incubator for 1 hour, and then proceed with plate laying.

Low Conductivity (0.080 ± 0.005 mS/cm): This is the most critical parameter for achieving high-efficiency electrofusion. A low-conductivity environment can minimize the harmful Joule heat generated during electrification and prevent cells from being burned. Simultaneously, it ensures that the electric field can more effectively act on the cell membrane at a given voltage, promoting cell alignment and perforation.

Specific Osmolarity (130-180 mOsm/kg): This osmolarity range creates a low osmotic environment, which helps cells to moderately absorb water and expand before the electrical pulse, making the cell membrane in a tensed state that is more easily punctured by electric shock, thereby further improving fusion efficiency.

This product is stable and can be safely transported at ambient temperature. Upon receipt, please store it at 2-8°C. The product has a shelf life of 18 months under the specified storage conditions. Please be sure to use the product before the expiration date indicated on the product label.