Your preparation had decomposed. Baló-Banga

@Joseph

Why did the prepration decompose??

hi..

The lack of elevation of TNF alpha, IL-6, and IL-1beta protein expression in your in-vitro study suggests that the LPS you are using may not be effective at inducing the inflammatory response you are trying to study.

Here are a few things you can try to evaluate the efficacy of your LPS:

It's important to consider the source of the tissue you are using in your in-vitro study. Different tissue types may have varying levels of responsiveness to LPS and other inflammatory stimuli. If possible, it might be helpful to compare the results of your LPS treatment on colon and small intestine tissues to another tissue type that is known to be responsive to LPS, such as primary macrophages or immune cells, as a positive control.

Also, different batches of animals or human sources may affect the results, as the baseline protein expression levels of the cytokines you are studying may vary. It would be ideal to compare the result with a positive control of the same tissue source, if possible.

It's also important to note that the inflammatory response to LPS can vary depending on the dose, duration, and timing of treatment. You might want to try different doses, incubation times and evaluate the protein expression levels at different time points after LPS treatment, to see if that changes the response.

In summary, there are several factors to consider when evaluating the efficacy of LPS, including purity, concentration, storage conditions, tissue source, and time and dose of treatment. It might be helpful to compare your results to a positive control and to evaluate the protein expression levels of your target cytokines at different time points after LPS treatment.

Very helpful, Thanks Dr Mohammad I will try inshallah