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Boster's ELISA Kit is for the detection of human HO-1 in cell lysates, tissue extracts, and serum samples. Each kit contains sufficient components to quantitate the HO-1 concentration in up to 40 samples, tested in duplicate.
| Product Name | HO-1 ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7112 |
| Description | Colorimetric detection of HO-1. 96wells/kit, with removable strips. |
| Cite This Product | HO-1 ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7112) |
| Validated Species | Human |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. **For positive and negative control design, consult "Tissue specificity" under Protein Target Info. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 0.21 ng/ml *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 0.781 - 50 ng/mL *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Cell lysates, Serum, Tissue *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store at 4°C. |
| Description | Quantity |
| Anti-HO-1 Immunoassay Plate | 1 Plate |
| 5X HO-1 Extraction Reagent | 1 vial/10 ml |
| Recombinant HO-1 Standard | 2 vials |
| Standard and Sample Diluent | 1 vial/ 50 ml |
| 10X Wash Buffer Concentrate | 1 vial/100 ml |
| Anti-HO-1 Biotinylated Antibody Concentrate | 1 vial/150 μl |
| Anti-HO-1 Biotinylated Antibody Diluent | 1 vial/ 13 ml |
| Streptavidin: HRP Concentrate | 1 vial/150 μl |
| Streptavidin: HRP Diluent | 1 vial/ 13 ml |
| TMB Substrate | 1 vial/ 13 ml |
| Stop Solution | 1 vial/ 13 ml |
| Pre-treatment Buffer | 1 vial/ 13 ml |
1. Ultra pure water.
2. Additional reagents and materials for cell lysate and tissue extract preparation, including protease inhibitors.
3. Precision pipettors, with disposable plastic tips.
4. Polypropylene or polyethylene tubes to prepare samples ? do not use polystyrene, polycarbonate or glass tubes.
5. A container to prepare 1X Wash Buffer.
6. A wash bottle or an automated 96-well plate washer.
7. Disposable reagent reservoirs.
8. A standard microtiter plate reader for measuring absorbance at 450 nm.
9. Adhesive plate sealers.
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | HMOX1 |
|---|---|
| Protein Name | Heme oxygenase 1 |
| Protein Function | Heme oxygenase cleaves the heme ring at the alpha methene bridge to form biliverdin. Biliverdin is subsequently converted to bilirubin by biliverdin reductase. Under physiological conditions, the activity of heme oxygenase is highest in the spleen, where senescent erythrocytes are sequestrated and destroyed. Exhibits cytoprotective effects since excess of free heme sensitizes cells to undergo apoptosis. |
| Tissue Specificity | Expressed at higher levels in renal cancer tissue than in normal tissue (at protein level). |
| Subcellular Localization | Microsome. |
| Uniprot ID | P09601 |
| Alternative Names | Heme oxygenase 1; HO-1 |
| Research Areas | Cancer, Oxidative Stress| |
*if product is indicated to react with multiple species, protein info is based on the human gene.
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 (6), 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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Typical Standard Curve for the HO-1 ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7112 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.781 – 50 ng/mL.
1. Prepare Standard and samples in Standard and Sample Diluent.
2. Add 50 ?L of Pre-Treatment Buffer to all sample and standard wells.
3. Add 50 ?L 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 ?L 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 ?L 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 ?L 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 ?L of Stop Solution to each well.
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