My Dear Friends,
1). I have A549 cell PM10 treated supernatant, i have to measure IL-1B, IL-6, IL-8 and TNF alpha influence after treatment. So i need protocol for this detection method without ELSIA KITs.
All ELSIA KITS are very expensive. please help me.
2). I need some of the reference articles for PM10 cytokine and chemokine activity.
3). Very less amount ELISA Kits.
4). Please give me suggestion for good ELISA.
Cytometric bead array could be cheaper than ELISA for you . Because you will realize 4 tests together.
Yes Ishak Ozel Tekin .., You are absolutely correct i have 5 industrial samples. so please help me
you can also try a fluorescently-labeled protein microarray (RayBiotech has some systems )
You have not explained what you are treating cells with?? BUT...If you suspect increased inflammation, you can first check for increased expression of transcription factors (such as NFkB and STAT3) in your cells by Western Blot. You can also add treated supernatant to a target cell and measure its levels of NFkB and STAT3. If NFkB and STAT3 levels do increase, then, your treatment maybe inducing IL's and TNF's. If you dont see increase of these NFkB and STAT3--then there is no need to do ELISA etc. for cytokines.
You can perform western blot if you have required antibody.
you can do PCR if primers can be managed, better to do real time PCR.
moreover Bead array and cytoflex are the other options.
The A549 cells you are treating may also produce nitric oxide (an early marker of inflammation). NO (nitric oxide) can be estimated by Griess assay which is cheap. But, standardization of Griess assay must be done carefully. Hope this helps!
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