I have developed an reagents for an IgM capture ELISA. Could someone assist me with a reference paper on how to determine the best dilution of conjugate antibody. I was advised to consider the dilution that gives an OD of 0.1 (negatives ) and an OD of 1.5- 2.0 (positive samples). Any alternative thoughts.
Hi,
Generally the approach is to prepare several dilutions of your target antibody and then look for the concentration that gives strong signal with low background which makes the optimum concentration. Have a look at the technical guide below, I hope it helps.
https://tools.thermofisher.com/content/sfs/brochures/TR0065-ELISA-guide.pdf
Moreover, I also assume the researchgate thread below to be relevant to your question:
https://www.researchgate.net/post/How_can_you_choose_the_optimal_dilution_of_antibodies_for_an_ELISA_assay
Dear Caroline Wasonga Otieno
Hello
Best way to test your own dilution
pls. find some references may help you
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC154952/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC164246/
http://www.mobitec.com/cms/products/bio/08_screening/rubella_virus_assays.html?pdf=ODZ-241.pdf
http://jcm.asm.org/content/37/4/1049.full
http://www.kpl.com/docs/techdocs/KPL%20ELISA%20Technical%20Guide.pdf
Regards
Prof. Houda KAWAS
Thank you Tareg Omer Mohammed and Prof. Houda KAWAS for the feedback.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA