I need do quantify the cytokines that are produced from dendritic cells descendant from monocytes when I infect these DCs with Trypanosoma cruzi in a 24-well-plate, but I have never worked with ELISA methods before. How ELISA kit is best for my aims?
Thank you.
The ELISA MAX kits from BioLegend are trustworthy. You will need to ensure you purchase the appropriate kit for the species of cells you are using (mouse, human, etc...). Also, by way of personal experience, you cannot purchase individual kit components to use in these kits. When you exhaust a reagent, you must order a complete new kit. These kits appear to be optimized to only work with the reagents included in the kit. Of note, you can purchase the plates for running these assays in bulk (Costar or Nunc), which may offer cost savings as well. I believe many of the kits for cytokines use the Costar plates.
Best of everything.
Jerome
It depends on your measuring cytokines and range and time. I suggest supersensitive FACS beads you can combine > 20 cytokines in one assay and time line is important for early an late response
BioLegend Llgendplex™, BD CBA,MACSPlex Cytokine
ELISA can do the job too
I have enough experience with the ELISA technique, but I would advise you to use another technique such as flow cytometry, I think that in your case, the what matters, is the quantification of intracellular citocines? ELISA is an option, but I hope you do your experiments by other techniques. I am not in favor of the kits, if I see an article based on it, I prefer not to read it.
Sometimes saving time is not the best option. It is only a personal recommendation.
It depends whether you will perform these assays regularly or only once. If only once, don't bother and buy a pre-coated assay. If not, DUOSET from R&D are very good and less expensive than pre-coated plates. It is 15 years I'm using them for human and mouse cytokines.
I suggest to use an ELISA kit from R & D systems. It has an attached protocol, which you can also find it online on their website, and it'll guide you through the procedure (step by step), but you have first to choose the suitable kit for the species of cells you are working on (mouse, human, etc...).
Dear Amanda,
From my own experience and if you want to measure MOUSE cytokines, I also can highly recommend the DuoSet ELISA Kits from R&D Systems. They might by slightly more expensive than the competitors kits, but I find them to work with superior accurarcy and reliability. Also, you can easily down-scale the reaction volume by half, i.e. from 100 to 50 ul/well. This way you safe reagents and make the kits more cost-effective. The down-scaling has worked for every single kit we tested (about 15 different ones). We also used NUNC high protein absorption plates.Good luck!
Tommy
I agree with the others suggestion. DuoSet ELISA Kits from R&D Systems or Raybiotech Kits are of good quality.
The DuoSet from R&D is a great option, not expensive and very sensible ELISA.
Dear Amanda,
I used in some experiments the BD Opteia kits, for assess the cytokines in supernatant of DC cells and T Cells. These kits have high specificity and sensibility.
I hope it can help you!!!
Douglas
What's the difference between ELISA Quantikine® kit ant Duoset ELISA kits?
Like most on here I use R & D kits, but I would advise using FACS Beads if you are looking at multiple cytokines. Once you identify the cytokines of interest you could use an appropriate ELISA kit.
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