Overview

  • Product name

  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 24 < 4%
    Inter-assay
    Sample n Mean SD CV%
    Overall 11 < 10%
  • Sample type

    Serum, Tissue Extracts, Cell Lysate, EDTA Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.036 ng/ml
  • Range

    0.195 ng/ml - 12.5 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Spike 101.5 96.7% - 107.91%

  • Assay time

    3h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Rat
    Does not react with: Mouse, Human
  • Product overview

    The rat HO-1 ELISA kit (ab213968) is a complete kit for the quantitative determination of rat HO-1 in cell lysates, tissue extracts, serum and plasma samples. It is recommended that you read the entire kit insert before proceeding with the assay.  The assay is specific for rat HO-1 and does not cross react with human or mouse HO-1.


    The HO-1 (rat), ELISA kit is a quantitative sandwich immunoassay. A mouse monoclonal antibody specific for HO-1 is pre-coated on the wells of the provided anti-rat HO-1 immunoassay plate.  HO-1 is captured by the immobilized antibody and is detected with a HO-1 specific, rabbit polyclonal antibody. The polyclonal antibody is subsequently bound by an anti-rabbit IgG antibody conjugated to horseradish peroxidase.


    The assay is developed with tetramethylbenzidine substrate (TMB) and a blue color develops in proportion to the amount of captured HO-1.  The color development is stopped with an acid stop solution which converts the endpoint color to yellow. The intensity of the color is measured in a microplate reader at 450nm. HO-1 concentrations from the sample are quantitated by interpolating absorbance readings from a standard curve generated with the calibrated HO-1 protein standard provided.

  • Notes

    Heme Oxygenase-1 (HO-1), also known as Hsp32, is the inducible isoform of heme oxygenase that catalyzes the NADPH, O2 and cytochrome P450 reductase dependent oxidation of heme to carbon monoxide, ferrous iron and biliverdin which is rapidly reduced to bilirubin.  These products of the HO reaction have important physiological effects: carbon monoxide is a potent vasodilator and has been implicated to be a physiological regulator of cGMP and vascular tone; biliverdin and its product bilirubin are potent antioxidants; “free” iron increases oxidative stress and regulates the expression of many mRNAs (e.g., DCT-1, ferritin and transferring receptor) by affecting the conformation of iron regulatory protein (IRP)-1 and its binding to iron regulatory elements (IREs) in the 5’- or 3’- UTRs of the mRNAs. 

    To date, three identified heme oxygenase isoforms are part of the HO system that catalyze heme into biliverdin and carbon monoxide.  These are inducible HO-1 or Hsp32, constitutive HO-2 that is abundant in the brain and testis, and HO-3, which is related to HO-2, however is the product of a different gene.  The HO system is the rate-limiting step in heme degradation and HO activity decreases the levels of heme which is a well-known potent catalyst of lipid peroxidation and oxygen radical formation.  The expression of HO-1 is highly responsive to all types of stimuli that cause oxidative stress and it is up-regulated during exposure to heat shock, oxidants, UV-A irradiation and other agents including cytokines, hormones, heme and heavy metals. 

    HO-1 is a vital component of neuronal defense mechanisms and oxidative stress has been postulated to be the underlying basis for neuronal cell death in neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease.  The expression of HO-1 is normally very low in the brain but increases markedly after heat shock, ischemia or glutathione depletion.  Spatial distribution of HO-1 expression in AD brain is essentially identical to that of the pathogenic conformational changes of tau protein that is the major component of the intraneuronal lesion of AD, neurofibrillary tangles. 

    HO-1 expression and tau expression may be regulated by oxidative stresses in a coordinated manner and play a pivotol role in the cytoprotection of neuronal cells.  Plasma and cerebrospinal fluid HO-1 protein and lymphocyte HO-1 mRNA levels are decreased in subjects with sporadic AD relative to normal elderly controls suggesting that measurement of HO-1 may serve as a useful biological marker in early sporadic AD.

    Oxidative stress in the heart caused by ischemia and reperfusion has been shown to lead to cardiomyocyte death.  An absence of HO-1 has detrimental consequences whereas overexpression of HO-1 plays a protective role in hypoperfusion and ischemia/reperfusion-induced myocardial injury.  Under normal conditions, HO-1 is present at low levels in all organs. The highest concentration of HO-1 can be found in testes, brain, and liver. However, expression in all organs is rapidly accelerated in response to pathophysiological conditions. Examples include renal ischemia/reperfusion and cellular transformation.  HO-1 overexpression exerts beneficial cytoprotective effects in a number of transplantation models, including antigen-independent ischemia/reperfusion injury, acute and chronic allograft rejection and xenotransplantation.

    The mechanisms by which HO-1 confers its protective effects are currently poorly understood but this area of investigation is active and rapidly evolving.  The measurement of HO-1 in various cell types, tissues and bodily fluids may provide new insights into the physiological roles of HO-1 and may lead to monitoring HO-1 levels as a biomarker for therapeutic interventions or as an environmental biomarker in toxicology studies.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab213968 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

Protocols

References

ab213968 has not yet been referenced specifically in any publications.

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