Human Cell Death ELISA Kits
Human HO1 (Heme Oxygenase 1) ELISA Kit (HUES03073)
- SKU:
- HUES03073
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P09601
- Sensitivity:
- 0.19ng/mL
- Range:
- 0.31-20ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- HO-1, HMOX1, HMOX1D, HSP32, Bk286b10
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Death
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.31-20 ng/mL |
| Sensitivity: | 0.19 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human HO1 in samples. No significant cross-reactivity or interference between Human HO1 and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human HO1. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human HO1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human HO1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human HO1. The concentration of Human HO1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | HMOX1: 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. Heme oxygenase 1 activity is highly inducible by its substrate heme and by various non-heme substances such as heavy metals, bromobenzene, endotoxin, oxidizing agents and UVA. Expressed at higher levels in renal cancer tissue than in normal tissue. Belongs to the heme oxygenase family. |
| UniProt Protein Details: | Protein type:EC 1. 14. 99. 3; Oxidoreductase; Cofactor and Vitamin Metabolism - porphyrin and chlorophyll Chromosomal Location of Human Ortholog: 22q13. 1 Cellular Component: caveola; cytosol; endoplasmic reticulum; endoplasmic reticulum membrane; extracellular space; membrane; nucleolus; nucleus; perinuclear region of cytoplasm Molecular Function:enzyme binding; heme binding; heme oxygenase (decyclizing) activity; metal ion binding; phospholipase D activity; protein binding; protein homodimerization activity; signal transducer activity Biological Process: angiogenesis; cell death; cellular iron ion homeostasis; cellular response to nutrient; DNA damage response, signal transduction resulting in induction of apoptosis; endothelial cell proliferation; erythrocyte homeostasis; excretion; healing during inflammatory response; heme catabolic process; heme oxidation; iron ion homeostasis; negative regulation of DNA binding; negative regulation of leukocyte migration; negative regulation of mast cell cytokine production; negative regulation of mast cell degranulation; negative regulation of neuron apoptosis; negative regulation of smooth muscle cell proliferation; negative regulation of transcription factor activity; porphyrin metabolic process; positive regulation of angiogenesis; positive regulation of chemokine biosynthetic process; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of smooth muscle cell proliferation; positive regulation of vasodilation; protein homooligomerization; regulation of angiogenesis; regulation of blood pressure; regulation of transcription factor activity; regulation of transcription from RNA polymerase II promoter in response to oxidative stress; response to estrogen stimulus; response to hydrogen peroxide; response to nicotine; response to oxidative stress; small GTPase mediated signal transduction; smooth muscle hyperplasia; transmembrane transport Disease: Heme Oxygenase 1 Deficiency; Pulmonary Disease, Chronic Obstructive |
| NCBI Summary: | Heme oxygenase, an essential enzyme in heme catabolism, cleaves heme to form biliverdin, which is subsequently converted to bilirubin by biliverdin reductase, and carbon monoxide, a putative neurotransmitter. Heme oxygenase activity is induced by its substrate heme and by various nonheme substances. Heme oxygenase occurs as 2 isozymes, an inducible heme oxygenase-1 and a constitutive heme oxygenase-2. HMOX1 and HMOX2 belong to the heme oxygenase family. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P09601 |
| NCBI GenInfo Identifier: | 123446 |
| NCBI Gene ID: | 3162 |
| NCBI Accession: | P09601. 1 |
| UniProt Related Accession: | P09601 |
| Molecular Weight: | 32,819 Da |
| NCBI Full Name: | Heme oxygenase 1 |
| NCBI Synonym Full Names: | heme oxygenase 1 |
| NCBI Official Symbol: | HMOX1 |
| NCBI Official Synonym Symbols: | HO-1; HSP32; HMOX1D; bK286B10 |
| NCBI Protein Information: | heme oxygenase 1 |
| UniProt Protein Name: | Heme oxygenase 1 |
| UniProt Gene Name: | HMOX1 |
| UniProt Entry Name: | HMOX1_HUMAN |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 20 | 2.364 2.374 | 2.369 | 2.292 |
| 10 | 1.519 1.531 | 1.525 | 1.448 |
| 5 | 0.83 0.818 | 0.824 | 0.747 |
| 2.5 | 0.433 0.451 | 0.442 | 0.365 |
| 1.25 | 0.258 0.254 | 0.256 | 0.179 |
| 0.63 | 0.184 0.154 | 0.169 | 0.092 |
| 0.31 | 0.124 0.124 | 0.124 | 0.047 |
| 0 | 0.074 0.08 | 0.077 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human HO1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human HO1 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 1.09 | 3.15 | 9.57 | 0.99 | 3.28 | 9.54 |
| Standard deviation | 0.07 | 0.18 | 0.39 | 0.05 | 0.15 | 0.48 |
| C V (%) | 6.42 | 5.71 | 4.08 | 5.05 | 4.57 | 5.03 |
Recovery
The recovery of Human HO1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 94-107 | 101 |
| EDTA plasma (n=5) | 92-110 | 100 |
| Cell culture media (n=5) | 84-97 | 91 |
Linearity
Samples were spiked with high concentrations of Human HO1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 89-103 | 99-111 | 88-103 |
| Average (%) | 95 | 105 | 94 | |
| 1:4 | Range (%) | 94-106 | 80-91 | 89-100 |
| Average (%) | 100 | 86 | 95 | |
| 1:8 | Range (%) | 88-100 | 85-101 | 86-97 |
| Average (%) | 94 | 92 | 92 | |
| 1:16 | Range (%) | 90-106 | 86-97 | 80-94 |
| Average (%) | 97 | 92 | 87 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
Related Products
Mouse HO1 (Heme Oxygenase 1) ELISA Kit
Rat HO1 (Heme Oxygenase 1) ELISA Kit
