Welcome friends to leave cryo rack in the comment area. So if the molecular weight of the two proteins you are studying is relatively large (such as GAPDH 37KD, Ki67 358KD), you can consider separating the glue from the middle, and the two parts are transferred at different times, which can achieve your desired effect. For some proteins with high expression peaks and stable expression, you can choose domestic production.
Cisco Jet provides two ultra-sensitive and ultra-sensitive ECL luminescent liquids for everyone to choose from. Dry texture of NC membrane is easy to brittle, mechanical strength and high solvent Tolerance No operation procedure Wet buffer solution, avoid air bubbles 100% methanol wetting test method before use ECL test ECL test is cheaper and more expensive Choice of film transfer method Wet transfer: Usually when we do wet transfer,
we choose 100V constant voltage (high strength, because the low strength time is longer, and the efficiency is lower), the current is controlled between 120-350mA, the molecular weight is below 60KD for 60 minutes, and the molecular weight above 60KD needs to extend the film transfer time 60 -150 minutes to ensure efficient transfer. Bonded to the surface. The necessary steps for the partners before loading are to determine the protein concentration and determine the protein concentration based on the protein concentration. According to laboratory habits and needs, friends can choose the method of membrane transfer. Although the principle of the Western Blot experiment is relatively simple, the experiments take a long time, the steps are tedious, and the experimental results are poorly reproducible.
The basic part of Western Blot is mentioned here first. Transfer membrane methods mainly include wet electroblotting (the best transfer membrane method), Semi-dry electroblotting (optional alternative membrane transfer method) and dry electroblotting (transmitting membrane method with limited sensitivity).
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