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Human Heme Oxygenase kit HTRF®

The Human Heme Oxygenase kit is designed for the rapid detection of HO-1 in cell lysates.
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  • Ready-to-use Ready-to-use
  • Highly specific Highly specific
  • Faster and more convenient than ELISA Faster and more convenient than ELISA
The Human Heme Oxygenase kit is designed for the rapid detection of HO-1 in cell lysates.
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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 kit enables the fast monitoring of HO-1 produced by cells.

Benefits

  • VALIDATED ON HUMAN LIVER CELLS
  • HO-1 MODULATOR STUDIES

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-1 kit principle

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.

protocol of the Human Heme Oxygenase kit

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.

Standard curve of the 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.

Induction of HO-1 in HEPG2 cells

Product Insert Heme Oxy (h) Kit / 63ADK044PEG-63ADK044PEH

63ADK044PEG-63ADK044PEH - Product Insert

HTRF Product Catalog 2020 July update

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Product Insert Human HOX-1 Kit / 64HOX1PEG-64HOX1PEH

64HOX1PEG-64HOX1PEH - Product Insert

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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