The NWLSS™ Human HO-1 ELISA kit provides a simple method to detect and quantify Heme Oxygenase (HO-1) in samples of human origin. The assay can be used with plasma, cell lysates and tissue homogenates. The assay does not cross react with the other known HO isoforms, HO-2 or HO-3
Heme Oxygenase-1 (HO-1) also known as Hsp32, is the inducible isoform of heme oxygenase that catalyzes NADPH, O2 and Cytochrome P450 reductase dependent oxidation of heme to carbon monoxide, ferrous iron and biliverdin. These products are involved in vasodilation, vascular tone and redox regulation. “Free” iron can increase oxidative stress and regulate the expression of many mRNAs by affecting the conformation of iron regulatory protein (IRP-1) and subsequent binding to iron regulatory elements (IREs). Three HO isoforms catalyzing heme into biliverdin and carbon monoxide have been identified: Inducible HO-1 (Hsp32), constitutive HO-2 abundant in brain and testis and HO-3 which is related to HO-2 but not of the same gene origin. HO activity decreases the levels of heme which is a catalyst for lipid peroxidation and oxygen radical formation. Expression of HO-1 is responsive to all types of oxidative stress related stimuli and it is up-regulated during exposure to oxidants, UV-A irradiation and a series of agents including heme, cytokines, hormones and heavy metals. Oxidative stress has been identified as a potential cornerstone in relation to neurodegenerative diseases such as Alzheimer’s (AD), Parkinson’s and ALS. HO-1 has been shown to play a role in neuronal defense of oxidative stress related events including heat shock and ischemia. Studies have shown that normal expression of HO-1 in the brain is typically low but increases after a heat shock or ischemic event. Additionally, spatial distribution of HO-1 in AD brains correlates with pathogenic changes in tau proteins associated with neurofibrillary tangles, a hallmark of AD brain lesions.
The Human HO-1 ELISA kit is a quantitative sandwich immunoassay. Murine monoclonal antibody specific for HO-1 is pre-coated on the wells of the plate provided. Sample and Standard HO-1 is captured by the stationary plate bound antibody. Captured HO-1 is then reacted with a polyclonal antibody specific for HO-1. The bound antibody-HO-1 complex is detected using an HRP -TMB system. First, anti-rabbit IgG antibody conjugated to horseradish peroxidase (HRP Conjugate) is added and allowed to react. After washing, tetramethylbenzidine (TMB) substrate is then added resulting in blue color development proportional to the amount of HO-1 present in each well. Color development is stopped using an acid stop solution changing the color to yellow which is measured and recorded at 450 nm. HO-1 concentrations in samples are measured by comparing sample 450 nm OD readings with the standard curve.
96 well sandwich ELISA
Quantitative measurement of human heme xxygenase-1 ( HO-1 or HSP32) in biological samples.
Human plasma, cell lystaes and tissue homogenates
Human Heme Oxygenase-1 (HO-1)
|Storage and Stability:||
1 year from manufacture date when components stored at specified temperatures
Microwells precoated with Anti Human HO-1|
Recombinant Human HO-1 Standard
500X Secondary Antibody
Secondary Antibody Diluent
500X HRP Reagent (Anti-rabbit IgG-HRP Conjugate)
20X Wash Buffer
5X Extraction Reagent