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Iron and Heme Core

Salt Lake City, Utah, US

The Iron and Heme Core’s primary mission is to facilitate research into the role of heme, heme precursors and transition metals in both normal and disease states. The iron and heme core lab has extensive experience with the separation and identification of tetrapyrroles and with running and developing heme biosynthesis pathway enzyme assays.

The Iron and Heme Core provides analysis of metals, precursor porphyrins and heme. The core will measure activity of the enzymes responsible for heme biosynthesis. Analysis and quantification of heme and its precursors can be obtained for cell pellets, tissue, whole blood, urine, feces and other complex biological materials. Analysis of enzyme activity can be provided for cell pellets, tissue and blood.

Selected Publications

  • Bergonia HA, Franklin MR, Kushner JP, Phillips JD. A method for determining δ´-aminolevulinic acid synthase activity in homogenized cells and... Show more »

The Iron and Heme Core’s primary mission is to facilitate research into the role of heme, heme precursors and transition metals in both normal and disease states. The iron and heme core lab has extensive experience with the separation and identification of tetrapyrroles and with running and developing heme biosynthesis pathway enzyme assays.

The Iron and Heme Core provides analysis of metals, precursor porphyrins and heme. The core will measure activity of the enzymes responsible for heme biosynthesis. Analysis and quantification of heme and its precursors can be obtained for cell pellets, tissue, whole blood, urine, feces and other complex biological materials. Analysis of enzyme activity can be provided for cell pellets, tissue and blood.

Selected Publications

  • Bergonia HA, Franklin MR, Kushner JP, Phillips JD. A method for determining δ´-aminolevulinic acid synthase activity in homogenized cells and tissues. Clin Biochem. 2015; 48(12):788-95.
  • Phillips JD, Kushner JP, Bergonia HA, Franklin MR. Uroporphyria in the
    Cyp1a2-/- mouse. Blood Cells Mol Dis. 2011; 47(4):249-54.
  • Warby CA, Phillips JD, Bergonia HA, Whitby FG, Hill CP, Kushner JP. Structural and kinetic characterization of mutant human uroporphyrinogen decarboxylases. Cell Mol Biol (Noisy-le-grand). 2009; 55(2):40-5.
  • Bergonia HA, Phillips JD, Kushner JP. Reduction of porphyrins to
    porphyrinogens with palladium on carbon. Anal Biochem. 2009; 384(1):74-8.
  • Phillips JD, Bergonia HA, Reilly CA, Franklin MR, Kushner JP. A porphomethene inhibitor of uroporphyrinogen decarboxylase causes porphyria cutanea tarda. Proc Natl Acad Sci U S A. 2007; 104(12):5079-84.
  • Yu MK, Bergonia H, Szabo A, Phillips JD. Progressive disease in chronic
    lymphocytic leukemia is correlated with the DNA methylation index. Leuk Res. 2007; 31(6):773-7. PubMed PMID: 16970987.
  • Mathews MA, Schubert HL, Whitby FG, Alexander KJ, Schadick K, Bergonia HA, Phillips JD, Hill CP. Crystal structure of human uroporphyrinogen III synthase. EMBO J. 2001; 20(21):5832-9.
  • Bulaj ZJ, Franklin MR, Phillips JD, Miller KL, Bergonia HA, Ajioka RS, Griffen LM, Guinee DJ, Edwards CQ, Kushner JP. Transdermal estrogen replacement therapy in postmenopausal women previously treated for porphyria cutanea tarda. J Lab Clin Med. 2000; 136(6):482-8.
  • Kim YM, Bergonia HA, Müller C, Pitt BR, Watkins WD, Lancaster JR Jr. Loss and degradation of enzyme-bound heme induced by cellular nitric oxide synthesis. J Biol Chem. 1995; 270(11):5710-3.
  • Kim YM, Bergonia HA, Müller C, Pitt BR, Watkins WD, Lancaster JR Jr. Nitric oxide and intracellular heme. Adv Pharmacol. 1995; 34:277-91.
  • Jackson LK, Brooks HB, Osterman AL, Goldsmith EJ, Phillips MA. Altering the reaction specificity of eukaryotic ornithine decarboxylase. Biochemistry. 2000; 39(37):11247-57.
  • Jackson LK, Brooks HB, Myers DP, Phillips MA. Ornithine decarboxylase promotes catalysis by binding the carboxylate in a buried pocket containing phenylalanine 397. Biochemistry. 2003; 42(10):2933-40.
  • Osterman AL(1), Brooks HB, Jackson L, Abbott JJ, Phillips MA. Lysine-69 plays a key role in catalysis by ornithine decarboxylase through acceleration of the Schiff base formation, decarboxylation, and product release steps. Biochemistry. 1999; 38(36):11814-26.
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UPLC
Ultra performance liquid chromatography
Price on request

Porphyrin Analysis

Porphyrins from urine, feces, cell lysates, serum, blood, media concentrates from mammalian cells and yeast and other biologic samples are analyzed using a Waters UPLC system. Porphyrins are separated on a reverse-phase C18 column, 2.1x 100 mm BEH, and visualized with the tandem high sensitivity photodiode... Show more »

Porphyrin Analysis

Porphyrins from urine, feces, cell lysates, serum, blood, media concentrates from mammalian cells and yeast and other biologic samples are analyzed using a Waters UPLC system. Porphyrins are separated on a reverse-phase C18 column, 2.1x 100 mm BEH, and visualized with the tandem high sensitivity photodiode array and fluorescence detectors of the UPLC.

Standard molecules analyzed include: heme (as hemin chloride), the precursor porphyrins (uroporphyrin, heptacarboxylporphyrin, hexacarboxylporphyrin, pentacarboxylporphyrin, and coproporphyrin) of both I and III isomers, protoporphyrin IX and zinc protoporphyrin IX. We may also assay for bilirubin and biliverdin. (Standards for most of these are available from Frontier Scientific, Logan, UT, and the remainder are generated by our lab)

There are instances where additional absorptive or fluorescent peaks are observed with “tetrapyrrole signatures” that do not correspond to the standard molecules listed above. When standards are not available and there is enough sample, fractions can be collected, pooled and analyzed by our sister CIHD core, the Metabolomics Core, at the University of Utah.

UPLC analysis of total heme and PPIX

A Waters Acquity ultra-high pressure liquid chromatography (UPLC) system equipped with a reverse-phase C18 column, a photodiode array detector and a fluorescence detector are used to determine total heme and PPIX. Samples are subjected to solvent extraction and all noncovalently associated heme and PPIX are solubilized and quantified. Hemin chloride (the Fe+3 equivalent of heme formed during sample processing) and PPIX are used as standards for quantitation.

Pricing includes sample processing, protein quantification for normalization (where needed), and assay run in triplicate.

Services Academic Pricing Commercial Pricing
Porphyrin Analysis by UPLC $37.53 per sample $49.71 per sample
UPLC analysis of total heme and PPIX $34.29 persample $45.42 per sample
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Protoporphyrin IX
Iron
HPLC
UPLC
Analytical HPLC
HPLC
UPLC
Urine
Biology
hematologic
hematology
serum
Anemia
Blood
Tissue
Cell Line
porphyria
enzyme assay
heme
porphyrin
coproporphyrin
feces
body fluid
feed
heme biosynthesis
heme
iron
erythropoiesis
ALAS
ALAD
PBGS
PBGD
U3S
UROD
COPOX
FECH
5-aminolevulinic acid synthase
aminolevulinate dehydratase
porphobilinogen synthase
porphobilinogen deaminase
uroporphyrinogen III synthase
uroporphyrinogen decarboxylase
coproporphyrinogen III oxidase
ferrochelatase
porphyrin
tetrapyrrole
heme biosynthesis
ALAS
PBGD
5-aminolevulinic acid synthase
aminolevulinate dehydratase
PBGS
porphobilinogen synthase
porphobilinogen deaminase
uroporphyrinogen III synthase
U3S
UIIIS
UROD
uroporphyrinogen decarboxylase
COPOX
coproporphyrinogen IX oxidase
FECH
ferrochelatase
iron metabolism
blood
hemoglobin
Enzyme Assays
Price on request

In addition to the price per sample (sample prep, protein quantification, assay in triplicate and analysis), the enzyme assays also have a one-time setup fee (for up to 12 samples for the ALAS assay and up to 25 samples for the other enzyme assays that we offer).

Pricing includes sample processing, protein quantification for... Show more »

In addition to the price per sample (sample prep, protein quantification, assay in triplicate and analysis), the enzyme assays also have a one-time setup fee (for up to 12 samples for the ALAS assay and up to 25 samples for the other enzyme assays that we offer).

Pricing includes sample processing, protein quantification for normalization, and assay run in triplicate.

Heme Biosynthetic Enzyme Assays Academic Pricing: (price per sample + setup fee) Commercial Pricing: (price per sample + setup fee)
ALAS (5-aminolevulinic acid synthase) $37.93 each + 64.04 $50.25 each + 84.82
ALAD (aminolevulinate dehydratase) $35.37 each + 18.68 $46.85 each + 27.74
PBGD (porphobilinogen deaminase) $35.11 each + 19.49 $46.51 each + 25.82
U3S (uroporphyrinogen III synthase) $36.55 each + 22.84 $48.41 each + 30.25
UROD (uroporphyrinogen III decarboxylase) $36.52 each + 18.98 $48.37 each + 25.14
COPOX (coproporphyrinogen III oxidase) $40.58 each + 27.50 $53.75 each + 36.43
FECH (ferrochelatase) $38.10 each + 22.07 $50.46 each + 29.23
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Protoporphyrin IX
HPLC
UPLC
Analytical HPLC
HPLC
UPLC
Waters UPLC
Biology
hematologic
hematology
Anemia
Blood
Tissue
Cell Line
porphyria
enzyme assay
heme
porphyrin
coproporphyrin
heme biosynthesis
heme
iron
erythropoiesis
ALAS
ALAD
PBGS
PBGD
U3S
UROD
COPOX
FECH
5-aminolevulinic acid synthase
aminolevulinate dehydratase
porphobilinogen synthase
porphobilinogen deaminase
uroporphyrinogen III synthase
uroporphyrinogen decarboxylase
coproporphyrinogen III oxidase
ferrochelatase
tetrapyrrole
heme biosynthesis
iron metabolism
5-aminolevulinic acid
hemin
heme metabolism
iron
blood
hemoglobin
Spectral Testing
Price on request

Spectral analysis of Heme

An Agilent 8453 diode array spectrophotometer can also be used for the spectrophotometric analysis of hemes. Typically, samples are solubilized in 200 mM NaOH in the presence of pyridine to obtain the pyridine-ferrohemochrome derivative spectra. Simultaneous determination of hemes a, b, and o, is... Show more »

Spectral analysis of Heme

An Agilent 8453 diode array spectrophotometer can also be used for the spectrophotometric analysis of hemes. Typically, samples are solubilized in 200 mM NaOH in the presence of pyridine to obtain the pyridine-ferrohemochrome derivative spectra. Simultaneous determination of hemes a, b, and o, is possible from the pyridine hemochrome spectra. Each heme species is quantified using the well-established alpha band extinction coefficients at characteristic absorbance maxima obtained from the reduced-minus-oxidized spectra.

Pricing includes sample processing, protein quantification for normalization (where needed), and assay run in triplicate.

Academic Pricing: $46.27
Commercial Pricing: $61.29

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Biology
hematologic
hematology
Anemia
porphyria
heme
spectrophotometer
spectrophotometry
pyridine
NaOH
Biology
Price on request
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Biochemistry & Molecular Biology
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Biochemical Assays
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Chemistry and Materials
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Analytical Chemistry Services
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Purification Services
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Chromatography
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Liquid Chromatography
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Computational Modeling
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Statistical Analysis
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