Forums

Question: The spots in 3 wells are blue

  • m
    mKim2024-09-26T00:19:56Z

    I measured the spots after the ELISpot using 3420-4HST-2 following the kit manual.

    ABC: Negative / DEF: Treated / GH: Positive

    1. The three untreated wells are blue. Could you explain the reason

    2. The number of spots differs from what is visible to the naked eye.

    If I decrease the sensitivity(intensity) and increase size, do you think the counts will align more closely with the visible numbers? Could you recommend how to adjust the reading conditions?"

    Let me know if you need any further adjustments!

    Picture1.jpg

  • C
    Christian@mabtech.com2024-09-26T06:33:20Z
    Admin

    Hi mKim!

    1. When using HRP in combination with TMB substrate from Mabtech you typically get a slight blue haze to the membrane. Spots are more distinct bluish. Totally normal. If you turn off the substrate reaction with water or PBS, this can change the color tone quite a lot depending on what you use. Should not have any impact on data.

    2. You ELISpot reader needs to be set up properly. What reader do you have? algorithm settings needs to be adjusted. Doing this will improve spot-counts to better match reality!