Signal Guard™ HRP reaction stopping reagent

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
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Physical properties

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Water

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

HRP coupling reactions provide sensitive biomolecular assays based on hydrogen peroxide- generating enzyme systems linked to peroxidase- mediated oxidation. Fluorogenic HRP substrates are preferred to use for enhancing assay sensitivities. Among them, the most commonly used HRP substrates include ADHP (also called Amplex® Red, #11000), Amplex® UltraRed and Amplite® Red. Typically, detection reactions are performed in microplate wells and are initiated by adding a fluorogenic HRP substrate, resulting in a continuous fluorescence increase. It is critical to ensure that the timing of the standard and unknown sample measurements is the same. Our Signal Guard™ HRP reaction stopping reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable. The Signal Guard™ HRP reaction stopping reagent is designed for use in conjunction with ADHP (Amplex® Red ), Amplite® and Amplex®UltraRed fluorogenic substrates. Under the same conditions, Signal Guard™ HRP reaction stopping reagent significantly outperforms the Amplex® Red/UltraRed Stop Reagent (#A33855) from ThermoFisher. Our Signal Guard™ HRP reaction stopping reagent can also be used in other HRP reaction systems.

Example protocol

AT A GLANCE

Important Note

Thaw each kit component at room temperature before starting the experiment.

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

Signal Guard™ HRP Reaction Stopping Reagent (20X)

Add 500 µL of ddH2O to Signal Guard™ HRP reaction stopping reagent vial and mix well.

Note Aliquot in small quantites and store at -20 °C in dark plac and avoid light.

PREPARATION OF WORKING SOLUTION

Signal Guard™ HRP Reaction Stopping Reagent (1X)

Add 50 µL of Signal Guard™ HRP reaction stopping reagent into 950 µL ddH₂O and mix well.
Note Prepare the working solutions freshly as needed, and avoid light.

SAMPLE EXPERIMENTAL PROTOCOL

At the desired stopping time point, add 20 µL of Stop Reagent solution (1X) per 100 µL volume in each microplate well.

Note: For other reaction volumes, adjust the addition of 1X Stop Reagent proportionally (e.g., add 5 µL to a 25 µL reaction volume). The 1X stop reagent should be added to all wells, including any reagent controls without HRP.

Note: The time-dependent fluorescence signal increase will terminate immediately, and the fluorescence signal level should remain stable for at least 5 hours.

Images

Figure 1. Application of HRP Reaction Stop Solution on HRP coupled glucose detection reaction. Two parallel reactions containing 15 µM Glucose were initiated by adding 50 µL assay mixture containing: 0.5mU/mL HRP, Amplex® Red, and 0.5mU/ml Glucose Oxidase. Reactions were incubated at room temperature for 5 mins and then 20 µL 1X Stop Reagent was added to one reaction, and 20 µL dd H2O to the other reaction. The plots demonstrated that the reaction is completely inhibited by Signal Guard™ HRP Reaction Stopping Solution.

Citations

View all 9 citations: Citation Explorer

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

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

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

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

Ordered cleavage of myeloperoxidase ester bonds releases active site heme leading to inactivation of myeloperoxidase by benzoic acid hydrazide analogs
Authors: Huang, Jiansheng and Smith, Forrest and Panizzi, Peter
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: Scherag, Frank D and Br, undefined and stetter, Thomas and Rühe, Jürgen
Journal: Colloids and Surfaces B: Biointerfaces (2014): 576--582

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

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

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

References

View all 61 references: Citation Explorer

Volatile fatty acid-sensing system involving coenzyme-A transferase
Authors: Rajashekhara E, Hosoda A, Sode K, Ikenaga H, Watanabe K.
Journal: Biotechnol Prog (2006): 334

Galactose oxidase action on galactose containing glycolipids--a fluorescence method
Authors: Fortelius M, Mattjus P.
Journal: Chem Phys Lipids (2006): 103

Effects of chronic ethanol administration on the activities and relative synthetic rates of myelin and synaptosomal plasma membrane-associated sialidase in the rat brain
Authors: Azuine MA, Patel SJ, Lakshman MR.
Journal: Neurochem Int (2006): 67

Mitochondrial reactive oxygen species in mice lacking superoxide dismutase 2: attenuation via antioxidant treatment
Authors: Morten KJ, Ackrell BA, Melov S.
Journal: J Biol Chem (2006): 3354

A new, sensitive method for enzyme kinetic studies of scarce glucosides
Authors: Mazura P, Fohlerova R, Brzobohaty B, Kiran NS, J and a L., undefined
Journal: J Biochem Biophys Methods (2006): 55

Evaluation of assays for the measurement of bovine neutrophil reactive oxygen species
Authors: Rinaldi M, Moroni P, Paape MJ, Bannerman DD.
Journal: Vet Immunol Immunopathol. (2006)

Kinetic analysis of semisynthetic peroxidase enzymes containing a covalent DNA-heme adduct as the cofactor
Authors: Fruk L, Muller J, Niemeyer CM.
Journal: Chemistry (2006): 7448

K+-independent actions of diazoxide question the role of inner membrane KATP channels in mitochondrial cytoprotective signaling
Authors: Drose S, Br and t U, Hanley PJ.
Journal: J Biol Chem (2006): 23733

A Role for Mitochondrial Dysfunction in Perpetuating Radiation-Induced Genomic Instability
Authors: Kim GJ, Fiskum GM, Morgan WF.
Journal: Cancer Res (2006): 10377

Sexual Dimorphism in Oxidant Status in Spontaneously Hypertensive Rats
Authors: Sullivan JC, Sasser JM, Pollock JS.
Journal: Am J Physiol Regul Integr Comp Physiol. (2006)