CBA024
InnoZyme Myeloperoxidase Activity Kit
Synonym(s):
InnoZyme MPO Activity Kit
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About This Item
UNSPSC Code:
41116133
NACRES:
NA.77
Recommended Products
usage
sufficient for 96 tests
Quality Level
species reactivity
human
packaging
pkg of 1 96-well plate(s)
manufacturer/tradename
Calbiochem®
storage condition
OK to freeze
avoid repeated freeze/thaw cycles
protect from light
assay range
sensitivity: 330 pg/mL
standard curve range: 5-100 ng/mL
input
sample type serum
sample type cerebrospinal fluid (CSF)
sample type cell lysate
detection method
colorimetric
shipped in
wet ice
storage temp.
−20°C
General description
The InnoZyme Myeloperoxidase Activity Kit is a specific and sensitive assay designed to measure myeloperoxidase activity and to screen enzyme inhibitors. The assay uses a polyclonal antibody specific for human MPO immobilized on a 96-well plate to specifically capture the enzyme. Activity of captured myeloperoxidase is measured using a detection reagent that includes TMB and hydrogen peroxide. Following color development, the reaction is stopped with sulfuric acid and the absorbance of the oxidized TMB is detected at 450 nm.
Components
Anti-MPO Coated 96-Well Plate, MPO Standard, TMB, Hydrogen Peroxide, Assay Buffer, Sample Buffer, Stop Solution, Plate Sealers, and a user protocol.
Warning
Toxicity: Multiple Toxicity Values, refer to MSDS (O)
Specifications
Assay Time: 2 h
Principle
The Calbiochem InnoZyme Myeloperoxidase Activity Kit is designed to measure human myeloperoxidase activity in cell lysates and biological samples and to screen enzyme inhibitors.
Preparation Note
All reagents necessary to perform the assay are supplied with the kit. Equilibrate all reagents to room temperature (15-25°C) just prior to use.Sample Buffer (1X): Dilute Sample Buffer 1:20 by adding 25 ml Sample Buffer to 475 ml dH2O. Myeloperoxidase Standard (1g/ml): Dilute the Myeloperoxidase Standard 1:100 by adding 5 l Myeloperoxidase Standard to 495 l chilled Sample Buffer (1X) obtain a concentration of 1 g/ml. Maintain the diluted Myeloperoxidase Standard (1 g/ml) on ice. Note: Following initial thaw the Myeloperoxidase Standard should be dispensed into aliquots and stored at -20°C. Avoid freeze/thaw cycles.<div class="Bio_doc_image">
Table 1: Myeloperoxidase Standard Dilutions
Prepare Myeloperoxidase Standard dilutions in the range of 5-100 ng/ml using the above guidelines.
</div>Hydrogen Peroxide (0.3%): Dilute the Hydrogen Peroxide (30%) 1:100 by adding 5 l Hydrogen Peroxide (30%) to 495 l chilled dH2O. Note: A total of 40 l is required to cover the entire Coated 96-Well Plate. If the entire plate is not to be used at one time, prepare fresh Hydrogen Peroxide (0.3%) each time the assay is performed. Following initial thaw the Hydrogen Peroxide should be dispensed into aliquots and stored at -20°C. Avoid freeze/thaw cycles.TMB Detection Reagent: Note: The desired amount of TMB Detection Reagent should be prepared fresh, just prior to use. To prepare TMB Detection Reagent, mix the following components:
Vortex gently 3 X 3 sTo prepare TMB Detection Reagent for a variable number of strips, use the following guidelines: <div class="Bio_doc_image">Table 2: Preparation of TMB Detection Reagent
To prepare TMB Detection Reagent for a variable number of strips, use the above guidelines.
</div>
Table 1: Myeloperoxidase Standard Dilutions
Prepare Myeloperoxidase Standard dilutions in the range of 5-100 ng/ml using the above guidelines.
</div>Hydrogen Peroxide (0.3%): Dilute the Hydrogen Peroxide (30%) 1:100 by adding 5 l Hydrogen Peroxide (30%) to 495 l chilled dH2O. Note: A total of 40 l is required to cover the entire Coated 96-Well Plate. If the entire plate is not to be used at one time, prepare fresh Hydrogen Peroxide (0.3%) each time the assay is performed. Following initial thaw the Hydrogen Peroxide should be dispensed into aliquots and stored at -20°C. Avoid freeze/thaw cycles.TMB Detection Reagent: Note: The desired amount of TMB Detection Reagent should be prepared fresh, just prior to use. To prepare TMB Detection Reagent, mix the following components:
- 7.16 ml dH2O
- 2.00 ml Assay Buffer
- 0.80 ml TMB
- 40 l H2O2(0.3%)
Vortex gently 3 X 3 sTo prepare TMB Detection Reagent for a variable number of strips, use the following guidelines: <div class="Bio_doc_image">Table 2: Preparation of TMB Detection Reagent
To prepare TMB Detection Reagent for a variable number of strips, use the above guidelines.
</div>
Dilute samples with Sample Buffer (1X) as needed. Note: If sample dilution is not required for sensitivity reasons, prepare the sample by adding 475 µl sample to 25 µl 20X Sample Buffer to ensure appropriate pH for the first incubation.
Storage and Stability
Upon arrival, store the unopened kit at -20°C. All kit components, once opened, can be stored for up to 3 months under the following conditions:
Store at 4°C: TMB, Anti-MPO Coated 96-Well Plate, Assay Buffer, Stop Solution, and Sample Buffer
Store at -20°C: Myeloperoxidase Standard, and Hydrogen Peroxide
Note: Following initial use the Myeloperoxidase Standard and the Hydrogen Peroxide should be dispensed into aliquots and stored at -20°C. Avoid freeze/thaw cycles.
Store at 4°C: TMB, Anti-MPO Coated 96-Well Plate, Assay Buffer, Stop Solution, and Sample Buffer
Store at -20°C: Myeloperoxidase Standard, and Hydrogen Peroxide
Note: Following initial use the Myeloperoxidase Standard and the Hydrogen Peroxide should be dispensed into aliquots and stored at -20°C. Avoid freeze/thaw cycles.
Analysis Note
Positive Control
Human myeloperoxidase
Human myeloperoxidase
Other Notes
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
Weil, S.C., et al. 1988. Science 240, 790.
Winterbourn, C.C., et al. 2000. Curr. Opin. Hematol. 7, 53.
Brennan, M.L., et al. 2003. New Engl. J Med. 349, 1595.
Zheng, L., et al. 2004. J. Biol. Chem. in press
Vita, J.A., et al. 2004. Circulation, 110, 1134.
Winterbourn, C.C., et al. 2000. Curr. Opin. Hematol. 7, 53.
Brennan, M.L., et al. 2003. New Engl. J Med. 349, 1595.
Zheng, L., et al. 2004. J. Biol. Chem. in press
Vita, J.A., et al. 2004. Circulation, 110, 1134.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Dam. 1 - Met. Corr. 1 - Skin Irrit. 2 - Skin Sens. 1
Storage Class Code
8A - Combustible corrosive hazardous materials
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Alessandro Trentini et al.
Inflammation research : official journal of the European Histamine Research Society ... [et al.], 63(5), 325-328 (2014-01-25)
We investigated the effect of balanced (BS) and unbalanced (UBS) solutions in the absence or presence of hydroxyethyl starch (HES) on neutrophil functionality, evaluating the release of matrix metalloproteinase (MMP)-9, myeloperoxidase (MPO), and MMP-8. Neutrophils were isolated by gradient centrifugation
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