QuantiChrom™ Heme Assay Kit

Protocol SDS

Application

• For quantitative determination of heme and evaluation of drug effects on heme metabolism.

Key Features

• Sensitive and accurate. Linear detection range 0.6 – 125 µM heme in 96-well plate assay.
• Simple and high-throughput. The “mix-and-read” procedure involves the addition of a single working reagent and reading the optical density. Can be readily automated as a high-throughput assay in 96-well plates for thousands of samples per day.
• Safety. Reagents are non-toxic.
• Versatility. Assays can be executed in 96-well plate or cuvet.

Method

• OD400nm

Samples

• Blood, serum, plasma, urine etc

Species

• All

Procedure

• 5 min

Size

• 250 tests

Detection Limit

• 0.6 µM

Shelf Life

• 12 months

More Details

Heme is one important member of the porphyrin family. It is synthesized in both mitochondria and cytoplasm and is a key prosthetic group for various essential proteins such as hemoglobin, cytochromes, catalase, and peroxidases. Heme determination is widely practiced by researchers of various blood diseases. Simple, direct, and automation-ready procedures for measuring heme concentration are becoming popular in Research and Drug Discovery. BioAssay Systems QuantiChrom™ Heme Assay Kit is based on an improved aqueous alkaline solution method, in which the heme is converted into a uniform colored form. The intensity of color, measured at 400 nm, is directly proportional to the heme concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples and exhibits high sensitivity.

Does the Heme Assay kit DIHM-250 detect heme bound to hemoglobin or only heme alone?

The heme assay detects the total heme content in the sample, including the heme in hemoglobin.

Does the heme assay detect β-hematin (malaria pigment) / protoporphyrin?

We have not specifically tested it, but it is highly likely that our heme assay will detect &beta:-hematin and protophorphyrin with the same efficiency as heme, and that it is not possible to distinguish between the variants.

Lapping-Carr, G., et al (2020). Circulating extracellular vesicles from patients with acute chest syndrome disrupt adherens junctions between endothelial cells. Pediatric Research. Assay: Heme in human plasma.

Li, H., et al (2020). SLC46A1 contributes to hepatic iron metabolism by importing heme in hepatocytes. Metabolism, 110, 154306. Assay: Heme in mouse serum.

Aggarwal, S., et al (2020). Heme attenuates endogenous opioid levels in leukocytes of HIV positive individuals with chronic widespread pain. Assay: Heme in human and mouse plasma.

Dos Santos, L. I., et al (2021). Disrupted iron metabolism and mortality during co-infection with malaria and an intestinal Gram-negative Extracellular pathogen. Cell Reports, 34(2), 108613. Assay: Heme in mouse plasma.

Keleku-Lukwete, N., et al (2019). Nrf2 activation in myeloid cells and endothelial cells differentially mitigates sickle cell disease pathology in mice. Blood Advances, 3(8), 1285-1297. Assay: Heme in mouse plasma.

Yang, X., et al (2020). Heme is involved in the systemic inflammatory response following radiofrequency ablation of hepatic hemangiomas. Assay: Heme in human serum.

Poillerat, V., et al (2020). Hemopexin as an inhibitor of hemolysis-induced complement activation. Frontiers in Immunology, 11. Assay: Heme in mouse plasma.

Koike, R., et al. (2020). Heat-killed Fusobacterium nucleatum triggers varying heme-related inflammatory and stress responses depending on primary human respiratory epithelial cell type. Molecules, 25(17), 3839. Assay: Heme in cell media.

Santaterra, V. A., et al (2020). Endothelial barrier integrity is disrupted in vitro by heme and by serum from sickle cell disease patients. Frontiers in Immunology, 11 Assay: Heme in human serum.

Aggarwal, S., Jilling, T., Doran, S., Ahmad, I., Eagen, J. E., Gu, S. & Patel, R. P. (2019). Phosgene Inhalation Causes Hemolysis and Acute Lung Injury. Toxicology Letters. 521724. Assay: Heme in mice blood.

Keleku-Lukwete, N., Suzuki, M., Panda, H., Otsuki, A., Katsuoka, F., Saito, R. & Yamamoto, M. (2019). Nrf2 activation in myeloid cells and endothelial cells differentially mitigates sickle cell disease pathology in mice. Blood advances, 3(8), 1285-1297. Assay: Heme in mice plasma.

Li, X., Wang, X., & Snyder, M. P. (2019). Metformin affects heme function as a possible mechanism of action. G3: Genes, Genomes, Genetics, 9(2), 513-522. Assay: Heme in yeast cells.

Belcher, J. D., Chen, C., Nguyen, J., Abdulla, F., Zhang, P., Nguyen, H. & Nath, K. A. (2018). Haptoglobin and hemopexin inhibit vaso-occlusion and inflammation in murine sickle cell disease: Role of heme oxygenase-1 induction. PloS one, 13(4), e0196455. Assay: Heme in mice plasma.

Cruz, L. A., Barral-Netto, M., & Andrade, B. B. (2018). Distinct inflammatory profile underlies pathological increases in creatinine levels associated with Plasmodium vivax malaria clinical severity. PLoS neglected tropical diseases, 12(3), e0006306. Assay: Heme in human blood.

Cueno, M. E., & Ochiai, K. (2018). Gingival periodontal disease (PD) level-butyric acid affects the systemic blood and brain organ: insights into the systemic inflammation of periodontal disease. Frontiers in immunology 9:1158. Assay: Heme in wistar rat blood.

Mooney, J. P., Barry, A., Goncalves, B. P., Tiono, A. B., Awandu, S. S., Grignard, L. & Riley, E. M. (2018). Haemolysis and haem oxygenase-1 induction during persistent “asymptomatic” malaria infection in Burkinabe children. Malaria journal, 17(1), 253. Assay: Heme in human plasma.

Santiago, R. P., Guarda, C. C., Figueiredo, C. V. B., Fiuza, L. M., Aleluia, M. M., Adanho, C. S. A. & Nascimento, V. M. L. (2018). Serum haptoglobin and hemopexin levels are depleted in pediatric sickle cell disease patients. Blood Cells Mol Dis. 72:34-36. Assay: Heme in human blood.

Abhishek, S., Gupta, A. K., & Singh, A. (2017). Kinetic Models Demonstrate Ability of Staphylococcus aureus to Uptake Heme from Beta vulgaris Proteins. Journal of Pure and Applied Microbiology, 11(4), 1713-1719. Assay: Heme in beetroot juice.

Dalko, E., Tchitchek, N., Pays, L., Herbert, F., Cazenave, P. A., Ravindran, B. & Pied, S. (2016). Erythropoietin levels increase during cerebral malaria and correlate with heme, interleukin-10 and tumor necrosis factor-alpha in India. PloS one, 11(7), e0158420. Assay: Heme in human plasma.

Luz, N. F., Balaji, S., Okuda, K., Barreto, A. S., Bertin, J., Gough, P. J. & Chan, F. K. M. (2016). RIPK1 and PGAM5 control Leishmania replication through distinct mechanisms. The Journal of Immunology, 196(12), 5056-5063. Assay: Heme in human plasma.

Ndisang JF, et al (2010). Up-regulating the heme oxygenase system with hemin improves insulin sensitivity and glucose metabolism in adult spontaneously hypertensive rats. Endocrinology 151(2):549-60. Assay: Heme in rat plasma.

Ndisang JF, Jadhav A (2010). The heme oxygenase system attenuates pancreatic lesions and improves insulin sensitivity and glucose metabolism in deoxycorticosterone acetate hypertension. American Journal of Physiolgy, Regululatory Integrative and Comparative Physiology. 298(1):R211-23. Assay: Heme in rat tissue.

Aldag C et al (2009). Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine. PNAS 106(14):5481-6. Assay: Heme in bacteria p450 enzymes.

Huang ML et al (2009). Elucidation of the mechanism of mitochondrial iron loading in Friedreich’s ataxia by analysis of a mouse mutant. PNAS 106(38):16381-6. Assay: Heme in mouse heart.

Zwart SR, et al (2009). Body iron stores and oxidative damage in humans increased during and after a 10- to 12-day undersea dive. J Nutr. 139(1):90-5. Assay: Heme in human plasma.

Pamplona, A. et al (2007). Heme oxygenase-1 and carbon monoxide suppress the pathogenesis of experimental cerebral malaria. Nature Med. 13(6): 703-710. Assay: Heme in mouse tissue.

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.