Amplite™ Blue

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240180120608642- Dose-ResponseData legend Generated with Quest Graph™ H2O2 (µM) RFU Hover mouse to interact
Hydrogen peroxide dose response was measured with Amplite™ Blue using a Gemini fluorescence microplate reader (Molecular Devices).
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Unit Size: Cat No: Price (USD): Qty:
25 mg 11005 $145


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Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
Email: sales@aatbio.com
International: See distributors





Overview

MW~300
SolventDMSO
Storage Freeze (<-15 °C)
Minimize light exposure
Category Enzyme Detection
Horseradish Peroxidase (HRP)
Related Secondary Reagents
A variety of substrates are available for detecting peroxidase activity in ELISA assays. Colorimetric HRP substrates (e.g., TMB, OPD and ABTS) have been widely used for years. Each of these substrates varies greatly with respect to its performance characteristics such as detection sensitivity, working range and attainable signal-to-noise ratios. Our Amplite™ Blue peroxidase substrate is a soluble fluorogenic substrate for detection of peroxidase activity. Amplite™ Blue allows rapid HRP detection assays to be performed with greater sensitivity than the colorimetric HRP substrates (such as TMB and OPD). The fluorescent product of Amplite™ Blue does not photobleach. Amplite™ Blue exhibits a flat baseline in assays, which facilitates low-level detection sensitivity and allows for high signal-to-noise ratios.




Calculators
Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Amplite™ Blue to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.



Molarity calculator

Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

Mass Molecular weight Volume Concentration Moles
/ = x =
 






Protocol


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This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

Important notes
It is important to store at <-15 °C and should be stored in cool, dark place.

It can be used within 12 months from the date of receipt.

Key parameters
Instrument:Fluorescence microplate reader
Excitation:324 nm
Emission:409 nm
Cutoff:350 nm
Recommended plate:Solid black
Example data analysis and figures

The reading (RFU) obtained from the blank standard well is used as a negative control. Subtract this value from the other standards' readings to obtain the base-line corrected values. Then, plot the standards' readings to obtain a standard curve and equation. This equation can be used to calculate H2O2 samples. We recommend using the Online Linear Regression Calculator which can be found at:

https://www.aatbio.com/tools/linear-logarithmic-semi-log-regression-online-calculator

Figure 1. Hydrogen peroxide dose response was measured with Amplite™ Blue using a Gemini fluorescence microplate reader (Molecular Devices).

Disclaimer
AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email info@aatbio.com if you have any questions.





References

Assessment of Tofacitinib and Ruxolitinib and their Anti Inflammatory Effects on Myeloperoxidase
Authors: Amber Milton
Journal: (2017)

Patterned Photonic Nitrocellulose for Pseudo-Paper ELISA
Authors: Junjie Chi, Bingbing Gao, Mi Sun, Fengling Zhang, Enben Su, Hong Liu, Zhongze Gu
Journal: Analytical Chemistry (2017)

Spinal Cord Inflammation: Molecular Imaging after Thoracic Aortic Ischemia Reperfusion Injury
Authors: Hassan Albadawi, John W Chen, Rahmi Oklu, Yue Wu, Gregory Wojtkiewicz, Benjamin Pulli, John D Milner, Richard P Cambria, Michael T Watkins
Journal: Radiology (2016): 152222

Myeloperoxidase Nuclear Imaging for Epileptogenesis
Authors: Yinian Zhang, Daniel P Seeburg, Benjamin Pulli, Gregory R Wojtkiewicz, Lionel Bure, Wendy Atkinson, Stefan Schob, Yoshiko Iwamoto, Muhammad Ali, Wei Zhang
Journal: Radiology (2015): 822--830

Myeloperoxidase--Hepatocyte--Stellate Cell Cross Talk Promotes Hepatocyte Injury and Fibrosis in Experimental Nonalcoholic Steatohepatitis
Authors: Benjamin Pulli, Muhammad Ali, Yoshiko Iwamoto, Matthias WG Zeller, Stefan Schob, Jenny J Linnoila, John W Chen
Journal: Antioxidants & redox signaling (2015): 1255--1269

Ordered cleavage of myeloperoxidase ester bonds releases active site heme leading to inactivation of myeloperoxidase by benzoic acid hydrazide analogs
Authors: Jiansheng Huang, Forrest Smith, Peter Panizzi
Journal: Archives of biochemistry and biophysics (2014): 74--85

Raising the shields: PCR in the presence of metallic surfaces protected by tailor-made coatings
Authors: Frank D Scherag, Thomas Brandstetter, Jürgen Rühe
Journal: Colloids and Surfaces B: Biointerfaces (2014): 576--582

Measuring myeloperoxidase activity in biological samples
Authors: Benjamin Pulli, Muhammad Ali, Reza Forghani, Stefan Schob, Kevin LC Hsieh, Gregory Wojtkiewicz, Jenny J Linnoila, John W Chen
Journal: PLoS One (2013): e67976

Micro-volume wall-less immunoassays using patterned planar plates
Authors: Katherine R Kozak, Jianyong Wang, Melvin Lye, Rashi Takkar, Namyong Kim, Hyunjae Lee, Noo Li Jeon, Kedan Lin, Crystal Zhang, Wai Lee T Wong
Journal: Lab on a Chip (2013): 1342--1350

Distinguishing inflammation from tumor and peritumoral edema by myeloperoxidase magnetic resonance imaging
Authors: Anne Kleijn, John W Chen, Jason S Buhrman, Gregory R Wojtkiewicz, Yoshiko Iwamoto, Martine L Lamfers, Anat O Stemmer-Rachamimov, Samuel D Rabkin, Ralph Weissleder, Robert L Martuza
Journal: Clinical Cancer Research (2011): 4484--4493






Documents

 
Safety Data Sheet (SDS)


Catalogs
1. Enzyme Probes & Assay Kits

Certificate of Analysis