I'm looking for a recommendation for an alternative to the Roche assay for analysis of 2,3-diphosphoglycerate (2,3-DPG) in blood, since they apparently have production issues with expected delay of at least a year.
The Roche assay is based on extraction through perchloric acid and subsequent neutralisation by potassium bicarbonate, and using this kit, 10 148 334 001, for detection by spectrophotometry (detection range of the kit is 0.02–0.15 umol).
I have freeze-stored samples already prepared according to the described method, awaiting analysis, but I also have blood units which I haven’t sampled yet. Ideally, I’d like to find a substitute to the kit, but changing method is also an option for my future samples.
I’d appreciate any recommendations very much!
Kind regards, Linda
Dear Linda Larsson, I just saw your question and I have the same problem. They are still not producing this kit and I am looking for an alternative way to measure 2,3-DPG in blood. Have you find any solution for yourself and would you like to share it?
Dear Linda and Fatih, We have the same problem and we are trying some antibody ELISA's at the moment. The main problem is that these ELISA's not work with extrated samples as we used in the Roche kit. Furthermore hemoglobin is a disturbing factor in these ELISA's. So far no results... :-(
If any of you finds a solution I hope you would share it with us. Best Regards from NL. Herbert
Dear Herbert,
I used this kit (https://www.mybiosource.com/23-bpg-general-elisa-kits/23-bisphosphoglycerate/288269) and we have some results. But I am not sure if it worked properly yet because the concentration of 2,3-DPG in RBCs was quite lower than expected. In this kit, they don't say anything about RBCs so I tried different dilutions and it was still very low. Maybe it is just not working, I cannot be sure for now. I need to try more samples but I don't have any experiments in the near future. Sorry for not being helpful at this moment.
Kind regards,
Fatih
Dear Fatih,
No problem, thanks for sharing. We had the same problem with this ELISA (mybiosource) very low values and also tried differnt dilutions. We tried another one (I have to look up the name and brand; working at home today) but this one worked with a standard diluent and a sample diluent and the sample diluent (without sample)already gave a huge signal (halfway the reference).
Best regards,
Herbert
Ran into this question when I started to look for a suitable assay for 2,3-DPG measurements. Is there a solution or a recommendation for another assay?
Dear Camilla, As far as we know, not yet. We have tried several 2,3-DPG ELISA kits but non of them works for our samples. Right now we are using a Hemox analyzer (very time consuming) to study the oxygen dissociation and association. Higher P50 values of the dissociation correlate with higher 2,3-DPG values. So at least we have an indication about 2,3-DPG levels. Best Regards Herbert
Has anyone by chance tried the procedure described here:
Poster Selective Recognition and Quantification of 2,3-bisphosphogl...
I am unfamiliar with the journal and none of the citations referencing back to the method it describes actually deployed it to my knowledge.
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