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HTRF Human Heme oxygenase-1 Detection Kit, 500 Assay Points

The Human Heme Oxygenase kit is designed for the rapid detection of HO-1 in cell lysates.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

Feature Specification
Application Protein Quantification
Sample Volume 16 µL

The Human Heme Oxygenase kit is designed for the rapid detection of HO-1 in cell lysates.

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

Product Variants
Unit Size: 500 Assay Points
Part #:
64HOX1PEG
List Price
USD 1,086.00
Unit Size: 10,000 Assay Points
Part #:
64HOX1PEH
List Price
USD 12,280.00

Overview

The human Heme Oxygenase 1 (HO-1) is an enzyme that catalyzes the degradation of Heme, leading to the production of biliverdin, ferrous iron, and carbon monoxide. HO-1 is an isoform induced in response to stress such as oxidative stress, hypoxia, heavy metals, cytokines, etc. The human Heme Oxygenase bles the fast monitoring of HO-1 produced by cells.

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
16 µL
Shipping Conditions
Shipped in Dry Ice
Target Class
Biomarkers
Target Species
Human
Technology
TR-FRET
Therapeutic Area
Cardiovascular
Infectious Diseases
Metabolism/Diabetes
Unit Size
500 Assay Points

Video gallery

How it works

Assay principle

Human heme oxygenase is measured using a sandwich immunoassay involving two specific human heme oxygenase antibodies, respectively labelled with Europium Cryptate (donor) and d2 (Acceptor). The intensity of the signal is proportional to the concentration of heme oxygenase present in the sample.

human-heme-oxygenase-kit

 

Assay Protocol

The human heme oxygenase kit features a two-plate assay protocol, where cells are plated, stimulated, and lysed in the same culture plate. Lysates are then transferred to the assay plate for the detection of Human HO-1. This protocol enables the cells' viability and confluence to be monitored. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.

human-heme-oxygenase-kit

 

Assay validation

Human Heme Oxygenase standard curve

A two-fold serial dilution starting from 25 ng/ml of Human Heme-Oxygenase-1 protein was performed, and fitted with with the 4 Parameter Logistic (4PL) model (with 1/Y² weighting). This data must not be substituted for the data obtained in the laboratory and should be considered only as an example.

human-heme-oxygenase-kit

 

Heme Oxygenase production in HEPG2 cells

HEPG2 cells were plated at 50,000 cells per well and were stimulated for 6H in the presence or absence of 10 µM of Cadmiumchloride. After removing the supernatant, the cells were lysed using the lysis buffer provided in the kit, and diluted 5 times before plating. The labeled antibodies from the kit were then added. The plate was read after 4 hours. A 2.9 fold increase of the HTRF ratio over non-stimulated cells was observed, exemplifying the production of human HO-1 after Cd-ion stimulation.

human-heme-oxygenase-kit

 

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