Inflammation is a vital biological process that protects and heals the body in response to harmful pathogens such as viruses, bacteria, and parasite. In some instances, inflammation goes into overdrive and becomes chronic, contributing to pathological conditions such as autoimmune diseases or facilitating cancer progression.

Cytokines play a central role in inflammation, either promoting or suppressing it. Their role has been studied extensively to understand the pathological inflammation mechanism, as potential therapies, or to assess response to treatment. Immunoassays are a standard tool for accurately measuring cytokine levels in a variety of biological samples.

Our cytokine immunoassays provide easy, consistent, and scalable ways to assess cytokine levels.

What do you need to consider when choosing an immunoassay for cytokine quantification?

If you need a ready-to-use kit, our SimpleStep ELISA kits® are 90-min single-wash ELISAs that are available in 96 or 384-well formats depending on the throughput.

You can also design your own immunoassay using our ready-to-conjugate antibody pairs

How do I select an ELISA kit?

For the cytokine you need to measure, you will need to consider:

• Is the kit validated for the right sample type

• The sensitivity and measuring range

• The sample volume needed

Our SimpleStep ELISA kits are validated in a broad selection of biological samples, including cerebrospinal fluid (CSF) for some. Their sensitivity and range have been validated to be relevant to the cytokine measured. The 384-well format (available for some of our kits) allows a higher throughput but also minimises sample volume requirements (1/4 volume compared to the 96-well format). Our kits are available for Human, mouse, and rat samples.

How do I select the best antibodies for my immunoassay

If you need to design your own immunoassay (bead-based for instance), you will need to find antibodies that have been validated as a pair in sandwich ELISA : these antibodies will recognise different epitopes on the target of interest and therefor can be used for sandwich-based immunoassay.

You will also need antibodies that can be conjugated easily and efficiently to the label of your choice.

Our BSA and azide-free (carrier-free) antibody pairs have been carefully selected for specificity, consistency, using recombinant monoclonal antibodies. Each pair consist of a capture and a detector antibody (best orientation in sandwich ELISA) available to purchase individually.

Proteins are essential in immunoassay development as they serve as both targets (antigens) and detection agents (antibodies), enabling specific recognition and quantification of analytes.

Finally, if you are considering multiplex immunoassays, you will need to make sure the antibody pairs you are using do not cross-react and can be used in the same assay. At Abcam, we have tested some of our pairs in immunoassay panels, and we have valuable cross-reactivity data that we can share on request.

For information about our pairs compatibility, please contact us.

References

1. Stefura, W. P., Graham, C., Lotoski, L., & HayGlass, K. T. Improved methods for quantifying human chemokine and cytokine biomarker responses: ultrasensitive ELISA and meso-scale electrochemiluminescence assays. Methods Mol. Biol. 2020, 91–114 (2019).
2. Chiswick, E. L., Duffy, E., Japp, B. & Remick, D. G. Detection and quantification of cytokines and other biomarkers. Methods Mol. Biol. 844, 15–30 (2012).
3. Lombardelli, L., Logiodice, F., Kullolli, O. & Piccinni, M.-P. Evaluation of secreted cytokines by multiplex bead-based assay (xMAP technology, Luminex). Methods Mol. Biol. 2285, 121–130 (2021).
4. Chaturvedi, A. K. et al. Evaluation of multiplexed cytokine and inflammation marker measurements: a methodologic study. Cancer Epidemiol. Biomarkers Prev. 20, 1902–1911 (2011).
5. Kany, S., Vollrath, J. T. & Relja, B. Cytokines in inflammatory disease. Int. J. Mol. Sci. 20, 6008 (2019).