Human TNF alpha ELISA Kit (ab100654)

Thank you for contacting The Scientific Support team.    Since this kit was not specifically validated with urine (only serum, plasma, and cell culture supernatants) unfortunately we do not have concentration levels to report.  In addition, when we tested normal serum with this kit, we found the measured levels were below the sensitivity limit which is 30 pg/ml (and not 15pg/ml as specified in the protocol, which will soon be changed to fix this mistake).  Therefore if testing body fluids, I would recommend using this kit mainly for spiking and recovery studies or for testing diseased or treated samples which have expected elevated TNF-alpha levels than normal. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us. We do not validate our ELISA kits with monkey samples, but I was able to find that human TNF-alpha shares 94 – 95% amino acid sequence homology with monkey. We do have a human TNF-alpha ELISA kit ab100654, which may likely work with your samples.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for taking the time to provide more information.

I am sorry to hear you have had difficulty obtaining satisfactory results from this kit.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab100654:

After reviewing the data I agree that the overall signals seem to be lower than expected, especially the standards. The most likely reason for this is that the Detection Antibody (Item F) was diluted with Assay Diluent A instead of Assay Diluent B.That's when we would see lower signals in tests.

I would also appreciate if you can confirm some further details:

What is the sample type?

At what temperature was the kit stored at and for how long?

Finally, this kit does not instruct the user to make a 666.7pg/ml or 74.07pg/ml standard.
The standard concentrations given in the kit are 6000, 2400, 960, 384, 153.6, 61.44, 24.58 and 0 pg/ml. Did you use a different dilution scheme?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

The equivalent IL10 assay, with 12 strips of 8 wells each, is ab100549. Product ab108871 comes with a single plate, not strips, but does use an HRP-conjugated streptavidin, as does the TNF alpha ELISA, ab100654.

Click here (or use the following: https://www.abcam.com/Human-IL-10-ELISA-Kit-ab100549.html).

Please contact us if you have any other questions.

Thank you for your interest in our IL6 and TNF alpha ELISA kits. My colleague will be happy to discuss pricing with you. As I mentioned, she will ask how many kits you want of each type, IL6 and TNF alpha. If you have any technical questions, our Scientific Support team is available from 8:30 in the morning to 8:00 in the evenening, EST.  

Thank you very much for contacting us. There is no fundamental difference between these kits. The ab46087 is based on sandwich ELISA procedure in which provided microtitre plate is coated with antibody. Ab100654 and ab108908 are based on direct ELISA procedure where antigens must be coated by incubating the plate with antigen or sample. We can assure that these KITS are equally suitable to detect TNF alpha in samples and are fully guaranteed by Abcam. I hope this information will be helpful. Please let us know if you have any questions or concerns.