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Conjugating biotinylated IL-21 with Strep-ALP Before Use in ELISpot?


Guest Philip Samaan

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Guest Philip Samaan

Hello all,

I am planning to conduct dual colour IL-21 and GzmB ELISpots in the future. In order to accomplish this, the detection antibody for IL-21 or GzmB needs to be conjugated with ALP. However, no such products are available in the market. The detection antibody for either cytokine is only found in the biotinylated form and the enzyme is sold separately as Strep-ALP. I'm wondering if it is possible to conjugate biotinlylated IL-21 detection antibody with Strep-ALP before use in ELISpot? Any help would be greatly appreciated.

Much thanks,

Philip Samaan

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Hello Philip and welcome to the forum!

It sounds to me like you are planning on doing a dual ELISpot with the enzymatic approach. We concluded long time ago that it simply does not work. 

One substrate will generate stronger responses than the other substrate and weak dual positive cells are simply identified as single secreting cells. Furthermore, the readers are rather horrible to use. 

Mabtech instead invested our time and resources into FluoroSpot. By coupling detection antibodies to fluorophores instead of enzymes correct analysis can be made. Also, with the launch of our own FluoroSpot reader IRIS in 2018, dual secreting cells are now recognized with accuracy for the first time due to the RAWspot algorithm. Even better, my own mother can run the machine it is that simple to use!

We can offer you IL-21/GzB FluoroSpot in our Flex format:

https://www.mabtech.com/products/flex/X-66B36W-1

 

Why not try a kit and send the plate for us for analysis!

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