Recombinant Human Fibronectin ,rFN

This protocol uses a cell culture coating experiment as an example and is intended for reference only. Actual application conditions may require optimization based on specific experimental needs ^[1].

1. Solution preparation:

Upon receipt, dissolve Recombinant Human Fibronectin ,rFN in sterile 1×PBS to prepare a stock solution at a concentration of 1mg/mL. Aliquot the stock solution into single-use volumes and store at –20°C to avoid repeated freeze-thaw cycles. Before use, thaw the required aliquot and dilute it with sterile 1×PBS (pH 7.2-7.4) to prepare the working solution. The recommended working concentration is 50-100μg/mL.

2. Plate coating (Example: 6-well plate)

Add approximately 800μL of the Recombinant Human Fibronectin ,rFN working solution to each well of a 6-well plate, ensuring that the entire bottom surface is evenly covered. Incubate the plate under one of the following conditions: overnight at 4°C, or for 1h at 37°C.

3. Working solution recovery

After incubation, collect the remaining Recombinant Human Fibronectin ,rFN working solution and store it at –20℃. It can be reused for coating other culture vessels within one week, provided sterility is maintained.

4. Cell Seeding

After coating, remove the residual solution if necessary. Add the appropriate volume of complete cell culture medium, and proceed with cell seeding directly. Recombinant Human Fibronectin ,rFN enhances cell adhesion, promotes cell spreading and migration, and supports optimal cell growth.

Notes:

1. If the Recombinant Human Fibronectin ,rFN working solution is not used immediately, it can be stored in a sealed container at –20℃. It is recommended to use it within one week to ensure optimal performance.

2. Adjust the volume of Recombinant Human Fibronectin ,rFN working solution according to the size and surface area of the culture plate or flask being used.

3. As a recombinant protein, Recombinant Human Fibronectin ,rFN should be handled gently. Avoid vigorous shaking during dissolution, and do not subject to repeated freeze-thaw cycles.

4. For laboratory safety, wear appropriate personal protective equipment such as a lab coat and disposable gloves during handling.

References:
[1]. Rappa G, Kunke D, Holter J, et al. Efficient expansion and gene transduction of mouse neural stem/progenitor cells on recombinant fibronectin[J]. Neuroscience, 2004, 124(4): 823-830.