JoVE Journal

Medicine

This content is Open Access.

High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease

Transcript

High throughput validation of multiple candidate biomarkers can be performed by sequential ELISA in order to minimize freeze/thaw cycles and use of precious plasma samples. Here, we demonstrate how to sequentially perform ELISAs for six different validated plasma biomarkers1-3 of graft-versus-host disease (GVHD)4 on the same plasma sample.

Chapters in this video

0:05

Title

1:34

Experiment Day 0: Sample Preparation and ELISA Test Plate Coating with Capture Antibody for IL-2Ralpha, REG3alpha and HGF

2:48

Experiment Day 1: IL-2Ralpha and REG3alpha ELISAs

4:13

Experiment Day 1: Elafin and TNFR1 Test Plate Coating with Capture Antibody

4:39

Experiment Day 1-2: HGF ELISA

5:46

Experiment Day 2: Elafin ELISA

6:16

Experiment Day 2: TNFR1 ELISA

6:45

Experiment Day 2: IL-8 ELISA

7:25

Results: Representative Optical Densities for the Standard Curve of 7 Different ELISA Plates Measuring REG3alpha Concentrations

8:06

Conclusion

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