Human Ultrasensitive TNFa ELISA Kit Has Good Quality and Detection Range

June 07, 2016

Boston Children's Hospital
Infectious Diseases
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen

Product Name:

Human Ultrasensitive TNFa ELISA Kit

Catalog Number:

KHC3014C

We are performing a TNFaLISA assay from Perkin-Elmer to confirm the preliminary hits of a high-throughput screening using human cells. We used this ultrasensitive TNFa ELISA kit from Invitrogen as a secondary confirmation assay method to detect TNFa production in supernatant human cells. We found that this ultra sensitive ELISA kit from Invitrogen works well and provides good quality data.

Experimental Design and Results Summary

Application

Detects TNFa production from human cells.

Starting Material

Human cells

Protocol Overview

As per the manufacture protocol (did not modify).1. Collect human blood and isolate desired cells from the blood 2. Incubate cells with stimuli for 24 hours at 37C degrees in 384-well plate 3. Collect supernatant from the plateThen use the ELISA Kit: 1. Add 50 uL incubation buffer per well 2. Add standard, control and sample supernatant to wells 3. Add 50 uL of Biotin Conjugate and incubate for 2 hours at 37C 4. Aspirate and wash 4x with wash buffer 5. Incubate 100 uL of Streptavidin-HRP Working Solution for 30 min at RT6. Aspirate and wash 4x with wash buffer7. Incubate 100 uL of Stabilized Chromogen for 30 min at RT8. Add 100 uL of Stop Solution and read plate at 450 nm (Total time: 3 hours)

Tips

The time before adding Stop Solution to stop the adding before reading the plate with a plate reader will be dependent on your sample concentrations and the reaction time needed

Results Summary

Good standard curve was achieved and the concentration of samples can be calculated.

Additional Notes

None

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Summary

The Good

Very easy to use the kit with all reagents provided and time efficient.

The Bad

None

The Bottom Line

I would recommend this TNFa ultrasensitive ELISA kit if you needed to detect low concentration of TNFa in your human samples. It is very easy to use, is a good quality assay kit and time efficient

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