Please how can cytokine result be used to obtain the gene expression of that particular cytokine such as interleukin 6.
Dear Emmanuel Ifeanyi Obeagu.
In fact, you can NOT convert protein expression data from ELISA experiments to mRNA expression data. Therefore, you have to perform additional experiments, e.g. RNA isolation, rev. transcription to cDNA and following qRT-PCR. By this way, you can get gene expression data (on mRNA level). After that, you can compare the mRNA expression data with your protein expression data (from performed ELISAs). And this will give you information of gene expression (e.g. of IL-6) and the following translation to the resulting protein.
In general:
1) Changes on mRNA level can be often detected earlier than on protein level. Depending on your target and stimulus mRNA changes can be possibly found at 8-24 h and resulting protein expression could then be analysed at 24-48 h (in an ideal manner you should perform expression analyses at different time point, to obtain information whether there is a linear or dynamic change and/or when expression change is max. and/or min.).
2) Sometimes mRNA and protein changes may follow the same direction (induced/reduced/not affected). However, it is also possible that induced mRNA expression is followed by reduced protein expression (i.e. via overload of the translational and/or splice apparature), and vice versa. Thus, it is necessary to do mRNA analyses (e.g. via qRT-PCR, gene expression arrays/chips or NGS) AND protein expression analyses (e.g. via Western blotting or ELISA or protein arrays).
I hope this is an adequate answer to your question and will help you!?
Sincerely
I mean how can you convert ELISA cytokine results to gene expression of the cytokines studied.
Dear Emmanuel Ifeanyi Obeagu.
In fact, you can NOT convert protein expression data from ELISA experiments to mRNA expression data. Therefore, you have to perform additional experiments, e.g. RNA isolation, rev. transcription to cDNA and following qRT-PCR. By this way, you can get gene expression data (on mRNA level). After that, you can compare the mRNA expression data with your protein expression data (from performed ELISAs). And this will give you information of gene expression (e.g. of IL-6) and the following translation to the resulting protein.
In general:
1) Changes on mRNA level can be often detected earlier than on protein level. Depending on your target and stimulus mRNA changes can be possibly found at 8-24 h and resulting protein expression could then be analysed at 24-48 h (in an ideal manner you should perform expression analyses at different time point, to obtain information whether there is a linear or dynamic change and/or when expression change is max. and/or min.).
2) Sometimes mRNA and protein changes may follow the same direction (induced/reduced/not affected). However, it is also possible that induced mRNA expression is followed by reduced protein expression (i.e. via overload of the translational and/or splice apparature), and vice versa. Thus, it is necessary to do mRNA analyses (e.g. via qRT-PCR, gene expression arrays/chips or NGS) AND protein expression analyses (e.g. via Western blotting or ELISA or protein arrays).
I hope this is an adequate answer to your question and will help you!?
Sincerely
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