Tips Shareing | Supertotal recombinant Protein Usage Guide

Recently, Abbkine has added a new batch of strictly verified active cytokine recombinant proteins to the shelves, many partners are consulting the use of recombinant protein precautions and preservation methods. Here it comes~

Abbkine recombinant proteins are freeze-dried powders that can be stored for several years at -20 °C or -80 °C. The lyophilized protein powder should be dissolved before use. Some recombinant protein products have a resolubilization buffer, and the matching resolubilization buffer is preferred. Resolubility and storage conditions of different recombinant proteins may be different. Refer to the recommended methods in the manual for details.

Dissolution methods

Always centrifuge the product before opening the tube. Centrifuge at 10,000-12000 rpm for 30 s or 3000-3500 rpm for 5 min. It is recommended that the lyophilized powder be redissolved in sterile endotoxin-free ddH₂O to 0.1-1mg /mL, during which time it can be gently blown and mixed with a gun, and left at room temperature for at least 20 min to completely dissolve. Be careful to avoid violent vortices.

How to use

It is recommended that the lyophilized powder be redissolved at a certain concentration in accordance with the instructions, and then diluted with a specific diluent according to the concentration required by the actual experiment.

How to store

lyophilized powder form: lyophilized powder form of recombinant protein is usually more stable, -20℃ or -80℃ can be preserved for a long time.

Liquid form after resolution: the redissolved recombinant protein can be stored for 1 week at 4℃.

Long-term preservation of complex solution:

Since neither the lyophilized solution of Abbkine recombinant protein nor the redissolved buffer contain protein protectants, if the dissolved protein solution needs to be preserved for a long time, it is recommended to add carrier protein (5% HSA, 10%FBS or 0.1%BSA) and store it in separate packages to avoid repeated freeze-thawing.

FAQ

Q: Why is it necessary to centrifugal before opening the cover?

A: The Abbkine recombinant protein does not contain carrier proteins or other additives (such as HSA, BSA or sucrose, mannitol, trehalose, etc.), and is usually freeze-dried with minimal salt solution and stored in a sterile tube. A trace amount of protein will be deposited in the tube wall during the lyophilization process, forming a thin or invisible protein film. The lyophilized powder may be dispersed and adhere to the tube wall or cap due to turbulence during transportation. Therefore, the lyophilized powder should be collected at the bottom of the tube before opening the cap to reduce the loss of lyophilized powder and ensure the correct concentration of dissolved protein.

Q: Why not simply dilute the protein to the concentration required for the experiment instead of redissolving it first?

A: The resolution buffer recommended in the instructions is the best condition for dissolving lyophilized powder, and direct dilution may cause insolubility. The protein can maintain good activity and stability in a certain concentration range. Below or above the concentration range, the protein may not be completely dissolved, and even the phenomenon of protein aggregation, or the activity is weakened or even lost.

Q: Why does freeze-dried powder sometimes not dissolve?

A: The solubility of the protein depends on many factors, such as pH and ionic strength, and the resolubilization buffers recommended in the instructions have been rigorously tested and are liquids that can completely dissolve the active protein. If it is difficult to dissolve immediately, it can be left at room temperature for 20 minutes to completely dissolve, or it can be placed on a horizontal shaker at a low speed for a period of time, or the suspension can be left at 4℃ for more than 2 hours.

Q: Can we use a vortex to shake violently and help the egg whites dissolve quickly?

A: Do not use vortexes for rapid oscillations. Generally, the recombinant protein can be completely dissolved with a few blows of the pipette's gun head. If it is not easy to dissolve, other factors should be considered and other methods should be tried to promote dissolution.

Q: Does the recombinant protein contain preservatives?

A: Abbkine recombinant proteins are sterile and preservative-free, and care should be taken to avoid microbial contamination during use. It is recommended to use sterilized solution to redissolve, use sterile tubes for sub-packaging and preservation, to avoid cross-contamination after repeated use.

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