Hsp 27 ELISA Kit

Introduction :

StressMarq Biosciences Inc.’s Molecular Signature™ Series ELISA Kit is for the detection of free, unbound human Hsp27 in cell lysates, tissue extracts, and human serum samples. The latter matrix has significant quantities of Hsp27-complexing components which can obscure the detection of Hsp27 by this assay. Each kit contains sufficient components to quantitate the Hsp27 concentration in up to 40 samples, tested in duplicate.

Assay Overview :

1. Prepare Standard and samples in Standard and Sample Diluent.

2. Add 100 μL of Standard or sample to appropriate wells.

3. Cover plate with Plate Sealer and incubate at room temperature (20-25°C) for 1 hour.

4. Wash plate four times with 1X Wash Buffer.

5. Add 100 μL of Biotinylated Antibody Working Solution to each well.

6. Cover plate with Plate Sealer and incubate at room temperature for 1 hour.

7. Wash plate four times with 1X Wash Buffer.

8. Add 100 μL of Streptavidin-HRP Working Solution to each well.

9. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.

10. Wash plate four times with 1X Wash Buffer.

11. Add 100 μL of TMB Substrate to each well.

12. Develop the plate in the dark at room temperature for 30 minutes.

13. Stop reaction by adding 100 μL of Stop Solution to each well.

14. Measure absorbance on a plate reader at 450 nm.

Assay Range :

13-0.2 ng/mL

• Suggested standard curve points are 13 ng/mL, 6.5 ng/mL, 3.25 ng/mL, 1.625 ng/mL, 0.813 ng/mL, 0.406 ng/mL, 0.203 ng/mL, and 0 ng/mL.

Assay Specificity and Species Reactivity :

• This assay is specific for Hsp27.

• The Hsp27 ELISA has been validated for the detection of free (unbound) human Hsp27.

Sensitivity :

• The lower limit of detection for the Hsp27 ELISA is 0.04 ng/mL.

Download datasheet