Cardiovascular and Oxidative Stress Assay Kit

HO-1 ELISA Assay Kit

$860.00 – $3,485.00

The HO-1 ELISA Assay Kit is for the detection of human HO-1 in cell lysates, tissue extracts, and serum samples.

SKU: SKT-111 Categories: , ,

HO-1 ELISA Assay Kit

The HO-1 ELISA Assay Kit is For Research Use Only

Sensitivity: 0.21 ng/ml
Dynamic Range: 0.781 – 50 ng/mL
Incubation Time: 30 minutes
Sample Type: Cell lysates,
Alternative Name: Heme-oxygenase 1

Product manufactured in Canada by StressMarq.

Assay Background to Heme-oxygenase 1 (HO-1)

Heme-oxygenase is a ubiquitous enzyme that catalyzes the initial and rate-limiting steps in heme catabolism yielding equimolar amounts of biliverdin, iron and carbon monoxide. Biliverdin is subsequently converted to bilirubin and the free iron is sequestered to ferritin. These products have important physiological effects as carbon monoxide is a potent vasodilator; biliverdin and bilirubin are potent antioxidants; and the free iron increases oxidative stress and regulates the expression of many mRNAs. There are three isoforms of heme-oxygenase, HO-1, HO-2 and HO-3; however HO-1 and HO-2 are the major isoforms as they both have been identified in mammals. HO-1, also known as heat shock protein 32, is an inducible isoform activated by most oxidative stress inducers, cytokines, inflammatory agents and heat shock. HO-2 is a constitutive isoform which is expressed under homeostatic conditions. HO-1 is also considered to be a cytoprotective factor in that free heme is highly reactive and cytotoxic, and secondly, carbon monoxide is a mediator inhibiting the inflammatory process and bilirubin is a scavenger for reactive oxygen, both of which are the end products of heme catalyzation. It has also been shown that HO-1 deficiency may cause reduced stress defense, a pro-inflammatory tendency, susceptibility to atherosclerotic lesion formation, endothelial cell injury, and growth retardation. Up-regulation of HO-1 is therefore said to be one of the major defense mechanisms of oxidative stress.

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Additional Information

Assay Procedure

  1. Prepare Standard and samples in Standard and Sample Diluent.
  2. Add 50 µL of Pre-Treatment Buffer to all sample and standard wells.
  3. Add 50 µL of Standard and sample to appropriate wells.
  4. Cover plate with Plate Sealer and incubate at room temperature (20-25°C) for 2 hours.
  5. Wash plate four times with 1X Wash Buffer.
  6. Add 100 µL of Biotinylated Antibody Working Solution to each well.
  7. Cover plate with Plate Sealer and incubate at room temperature for 1 hour.
  8. Wash plate four times with 1X Wash Buffer as described in step 5.
  9. Add 100 µL of Streptavidin-HRP Working Solution to each well.
  10. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.
  11. Wash plate four times with 1X Wash Buffer as described in step 5.
  12. Add 100 µL of TMB Substrate to each well.
  13. Develop the plate in the dark at room temperature for 30 minutes.
  14. Stop reaction by adding 100 µL of Stop Solution to each well.
  15. Measure absorbance on a plate reader at 450 nm.

Typical Standard Curve

HO-1 ELISA Assay Kit

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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