ELISpot Pro: Human IFN - γ (ALP) human interferon detection

ELISpot Pro: Human IFN - γ (ALP) human interferon detection

MABTECH ELISPOT experimental principle

The ELISPOT experiment used a PVDF membrane based 96 well plate (cat: MSIPS 4510, Millipore) coated with specific monoclonal antibodies (non-toxic, endotoxin free, high affinity antibodies are required) to capture cytokines secreted by cells.

Then add the cells to be detected and antigen stimuli into the wells of the culture plate for cultivation. Under the stimulation of stimuli, T cells will secrete corresponding cytokines during the corresponding time period. At this point, cytokines are captured by antibodies wrapped around the membrane.

After washing away the cells, the captured cytokines can bind to biotin labeled secondary antibodies, and then bind to biotin using enzyme-linked immunosorbent assay (ELISA) to form circular spots on the membrane (of course, fluorescein labeled secondary antibodies can also be used, which eliminates the need for subsequent chemical staining methods and can detect multiple cytokines at once). Each spot corresponds to a cell that initially secreted cytokines.

By counting the number of spots on the membrane and dividing it by the total number of cells added to the well, the percentage of positive cells can be calculated.

MABTECHThis technology has the following three major advantages:

1： High sensitivity. As long as one cell in a million secretes cytokines, they can be detected, which is 2-3 orders of magnitude higher than traditional ELISA methods.

2： Single cell level, live cell functional testing. ELISPOT directly stimulates live cells with antigens and detects the function of live cells.

3： Easy and economical operation, capable of high-throughput screening. ELISPOT does not have a complex in vitro cell expansion process, does not use isotopes, and according to standardized experimental procedures, one experimenter can process hundreds of samples simultaneously, with much higher efficiency than other detection methods.

ELISpot Pro: Human IFN - γ (ALP) human interferon detection

MABTECH: 3420-2APT-2 includes:

Pre-coated plates, mAb 1-D1K(2 clear plates) Detection mAb: 7-B6-1, ALP (150 μl)

BCIP/NBT-plus substrate(25 ml)

Positive control: Anti-CD3 mAb (CD3-2) (100 μl)

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