Mouse Heme Oxygenase 1 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Mouse Heme Oxygenase 1 with a sensitivity of 12 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Application | Reactivity | Dilution info | Notes |
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Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Heme oxygenase 1. Catalyzes the oxidative cleavage of heme at the alpha-methene bridge carbon, released as carbon monoxide (CO), to generate biliverdin IXalpha, while releasing the central heme iron chelate as ferrous iron (By similarity) Affords protection against programmed cell death and this cytoprotective effect relies on its ability to catabolize free heme and prevent it from sensitizing cells to undergo apoptosis (By similarity). Heme oxygenase 1 soluble form. Catalyzes the oxidative cleavage of heme at the alpha-methene bridge carbon, released as carbon monoxide (CO), to generate biliverdin IXalpha, while releasing the central heme iron chelate as ferrous iron.
Heme oxygenase 1, HO-1, P32 protein, Hmox1
Mouse Heme Oxygenase 1 ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Mouse Heme Oxygenase 1 with a sensitivity of 12 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Sample | n | C.V. |
---|---|---|
Sample Cell Media | n 5 | C.V. 2.4 |
Sample | n | C.V. |
---|---|---|
Sample Cell Media | n 3 | C.V. 2.6 |
Sample type | Average % | Range |
---|---|---|
Sample type Serum | Average % = 96 | Range 94 - 98 % |
Sample type EDTA Plasma | Average % = 89 | Range 87 - 91 % |
Sample type Urine | Average % = 105 | Range 102 - 111 % |
Sample type Tissue Extracts | Average % = 110 | Range 104 - 113 % |
Sample type Heparin Plasma | Average % = 88 | Range 86 - 91 % |
Sample type Citrate plasma | Average % = 95 | Range 87 - 108 % |
Sample type Cell culture media | Average % = 112 | Range 108 - 119 % |
Heme Oxygenase 1 (HO 1) in vitro CatchPoint® SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Heme Oxygenase 1 (HO 1) protein in mouse serum, plasma, urine, cell culture supernatant, and cell and tissue extract samples.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.
The CatchPoint® SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
Heme oxygenase 1 (HO 1) is an enzyme that functions in heme catabolism. The activity of heme oxygenase 1 is to cleave the heme ring to form biliverdin. There are two isozymes of heme oxygenase that have 47% amino acid sequence identity: inducible heme oxygenase 1 and constitutively expressed heme oxygenase 2.
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Heme Oxygenase 1 also known as HO-1 or HMOX1 is an enzyme that plays an important mechanistic role in heme catabolism. It catalyzes the degradation of heme into biliverdin carbon monoxide and free iron. This process involves the cleavage of the heme ring. HO-1 has a molecular weight of approximately 32 kDa. It is widely expressed in numerous tissues but is especially abundant in the liver and spleen. Its expression is induced by heme and other stress stimuli such as heavy metals cytokines and reactive oxygen species.
Heme Oxygenase 1 serves important protective functions in the body. It is not part of a larger complex but its products such as carbon monoxide and biliverdin have their own biological activities. Carbon monoxide produced by HO-1 has antiflammatory properties and can modulate apoptotic pathways. Biliverdin is reduced to bilirubin which acts as an antioxidant. The enzyme therefore directly influences cellular stress responses and maintains cellular homeostasis through these processes.
Heme Oxygenase 1 is integrally involved in oxidative stress response and heme metabolism. It participates in the cellular response to oxidative damage by reducing oxidative stress and promoting cytoprotection. Through its heme degradation activity it is connected with the synthesis of biologically active molecules like bilirubin and carbon monoxide. Heme Oxygenase 1 activity is related to other proteins in oxidative stress pathways such as Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2) which regulates its expression and globins which are sources of heme for HO-1 activity.
Heme Oxygenase 1 has been linked to conditions like cardiovascular diseases and neurodegenerative disorders. Its expression can attenuate the severity of atherosclerosis where oxidative stress is an important factor. In neurodegenerative diseases HO-1’s antioxidant properties may provide neuroprotection by mitigating oxidative damage. The protein's interactions with inflammatory cytokines such as Interleukin-6 and tumor necrosis factor-alpha influence its activity in these disease contexts.
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Interpolated concentrations of Heme Oxygenase 1 in mouse serum, and splenocyte supernatant.
The concentrations of Heme Oxygenase 1 were measured in duplicate and interpolated from the Heme Oxygenase 1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Heme Oxygenase 1 concentration was determined to be 10,670 pg/mL in mouse serum and 6,084 pg/mL in mouse splenocyte supernatant.
Interpolated concentrations of Heme Oxygenase 1 in mouse plasma (EDTA), plasma (heparin) and plasma (citrate).
The concentrations of Heme Oxygenase 1 were measured in duplicate and interpolated from the Heme Oxygenase 1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Heme Oxygenase 1 concentration was determined to be 5,536 pg/mL in mouse plasma (EDTA), 6,400 pg/mL in mouse plasma (heparin), and 5,479 pg/mL in mouse plasma (Citrate).
Quantitation of Heme Oxygenase 1 expression in different cell extracts.
Interpolated values of Heme Oxygenase 1 are plotted for the indicated cell extracts (y-axis) against the total extract load (x-axis).
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