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Recombinant Human HO-2/HMOX2 Protein, CF

Name: Recombinant Human HO-2/HMOX2 Protein, CF
Brand: Novus Biologicals
SKU: 3170-HM
Price: 423 USD

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Product Details

Summary

Reactivity	HuSpecies Glossary
Applications	Enzyme Activity
Format	Carrier-Free

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Recombinant Human HO-2/HMOX2 Protein, CF Summary

Details of Functionality	Measured by its ability to oxidize hemin to biliverdin. The specific activity is >3.5 pmol/min/μg, as measured under the described conditions.
Source	E. coli-derived human HO-2/HMOX2 protein Ser2-Leu291, with a C-terminal 6-His tag
Accession #	P30519
N-terminal Sequence	Ser2
Protein/Peptide Type	Recombinant Enzymes
Gene	HMOX2
Purity	>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Note	<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions	Enzyme Activity
Theoretical MW	34 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE	31 & 35 kDa, reducing conditions

Packaging, Storage & Formulations

Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

6 months from date of receipt, -70 °C as supplied.
3 months, -70 °C under sterile conditions after opening.

Buffer

Supplied as a 0.2 μm filtered solution in Tris and NaCl.

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Assay Procedure

Assay Buffer: 50 mM Tris, pH 7.5
Recombinant Human HO‑2/HMOX2 (rhHO‑2) (Catalog # 3170-HM)
Recombinant Human POR/Cytochrome P450 Reductase (rhPOR) (Catalog # 6340-PR)
Recombinant Human Biliverdin Reductase A/BLVRA (rhBLVRA) (Catalog # 6454-BR)
Catalase (Sigma, Catalog # C30), 100,000 U/mL stock
Hemin (Sigma, Catalog # H9039), 10 mM stock in DMSO
Bovine Serum Albumin (BSA), 100 mg/mL stock in deionized water
beta -Nicotinamide adenine 2'-phosphate reduced tetrasodium salt hydrate (NADPH) (Sigma, Catalog # N7505), 20 mM stock in deionized water
96-well Clear Plate (Costar, Catalog # 92592)
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

Dilute Hemin to 1 mM in Assay Buffer.
Prepare Reaction Mixture containing 60 µM Hemin, 4 mg/mL BSA, 1000 U/mL catalase, 80 µg/mL rhPOR, and 80 µg/mL rhBLVRA in Assay Buffer.
Prepare 80 µg/mL rhHO-2 in Assay Buffer.
Dilute NADPH to 1 mM in Assay Buffer.
In a clear plate, load 25 µL dilute rhHO-2 and add 25 µL Reaction Mixture. Include an enzyme blank containing 25 µL Assay Buffer and 25 µL Reaction Mixture.
Start the reaction by adding 50 µL 1 mM NADPH to all wells used.
Read absorbance at 468 nm (bottom read) in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (OD/min) x well volume (L) x 10¹² pmol/mol
	ext. coeff (M^-1cm^-1) x path corr.* (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Using the extinction coefficient 43500 M^-1cm^-1
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Reaction:

rhHO-2: 2 µg
rhPOR: 2 µg
rhBLVRA: 2 µg
Catalase: 250 U/mL
Hemin: 15 µM
BSA: 1 mg/mL
NADPH: 0.5 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human HO-2/HMOX2 Protein, CF

heme oxygenase (decycling) 2
heme oxygenase 2
HMOX2
HO2
HO-2
HO-2EC 1.14.99.3

Background

Heme oxygenase (HO) is the rate limiting enzyme in heme catabolism (1). It cleaves heme to biliverdin, carbon monoxide, and iron; biliverdin is subsequently converted by biliverdin reductase to bilirubin. The mechanism of HO is unique in that heme serves as both the substrate of the enzyme and as the prosthetic group for the activation of iron-bound O₂. HO activity is highest in spleen where senescent erythrocytes are sequestered and destroyed. Two isoforms, HO-1 and HO-2, are expressed in most tissues. HO-1 is an inducible enzyme in response to heavy metals, oxidative stress, cytokines, and many drugs (2). HO-2 is constitutively expressed in the brain and testes. HO-1 is expressed mainly in spleen, liver, and kidney. In addition to its activity in the heme degradation pathway, HO-2 potentially serves as an oxygen sensor through its heme regulating motifs, which have the core sequence of Cys-Pro (3).

Nader, G. (2008) Pharmacol. Rev. 60:79.
Otterbein, L.E. and A.M.K. Choi (2000) Am. J. Physiol. Lung Cell Mol. Physiol. 279:1029.
Yi. L. et al. (2009) J. Biol. Chem. 284:20556.

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Bioinformatics

Gene Symbol	HMOX2
Uniprot	P30519()

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Recombinant Human HO-2/HMOX2 Protein, CF

Species Glossary

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Product Details

Order Details

Recombinant Human HO-2/HMOX2 Protein, CF Summary

Applications/Dilutions

Packaging, Storage & Formulations

Notes

Alternate Names for Recombinant Human HO-2/HMOX2 Protein, CF

Background

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