We are inducing a periplasmic protein using 1mM IPTG at 37 degrees for 3 hours.
We cannot centrifuge and remove supernatant the same day.
So can we store the broth containing Bacterial cells and iptg for overnight at 4 degrees? And separate cells the next day.
You sure "Can", but results may vary. In some cases, IPTG induction is done at low temperatures (I used 18C) to allow for the expression of hard-to-fold proteins so that it might work. I suggest collecting small cell samples by microtube centrifugation and testing for protein expression by gel/western-blot.
Yes, it is generally acceptable to store an IPTG-induced culture broth at 4°C overnight. IPTG (isopropyl β-D-1-thiogalactopyranoside) is commonly used as an inducer in molecular biology experiments, particularly in the expression of recombinant proteins using IPTG-inducible promoters.
After inducing the culture with IPTG, you can store the culture broth at 4°C for a short period, typically overnight. The lower temperature helps slow down bacterial metabolism and protein degradation, allowing you to preserve the induced state of the cells.
Here are a few considerations when storing IPTG-induced culture broth at 4°C overnight:
1. Time Limit: Overnight storage generally refers to a period of approximately 12-16 hours. Storing the culture for longer periods may lead to changes in protein expression levels or reduced cell viability.
2. Sterility: Ensure that the culture broth is stored in a sterile container or properly sealed to prevent contamination during storage.
3. Agitation: If the culture broth is prone to settling or sedimentation, gently agitate or mix the broth before storage to ensure homogeneity when retrieving a sample the following day.
4. Monitoring: It is recommended to check the culture after overnight storage to assess the cell viability and evaluate the protein expression or any other desired outcomes. This assessment will help determine if the storage conditions and duration are appropriate for your specific experiment.
Remember that the optimal storage conditions may vary depending on the specific bacteria, expression system, and experimental requirements. Therefore, it is always advisable to consult the specific protocols or literature for your experimental setup to ensure the best results.
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