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ReadiUse™ Preactivated HRP NHS ester

Mechanism for conjugation of horseradish peroxidase (HRP) to target protein using activated NHS ester.
Mechanism for conjugation of horseradish peroxidase (HRP) to target protein using activated NHS ester.
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Catalog Number11025
Unit Size
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Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
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Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


ReadiUse™ Preactivated HRP NHS ester is the mono NHS ester of HRP. It can be used to readily label proteins (such as antibodies) or other biologicals molecules that have an amino group. ReadiUse™ Preactivated HRP NHS ester is robust and easy to use. It can be used to label antibodies with a simple mixing, i.e. the basic solution of an antibody (pH 8.5-9.0) can be directly mixed with the HRP NHS ester, and the reaction mixture is shaken for 1-2 hours. In most cases, the resulted solution can be directly used for ELISA assays without further purification. ReadiUse™ Preactivated HRP NHS ester provides the most robust method to perform the conjugation of antibodies with HRP. Any suitable amount of antibody (>1 mg/mL) can be readily conjugated to the preactivated HRP NHS ester without the need for purification.

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

  1. Antibody (IgG) ~100 µg in 20 µL~50µL of 25mM Sodium Bicarbonate Buffer (pH=8.5).

  2. Transfer IgG solution into HRP-SE vial, mix well.

  3. Reaction for 1~2 hours at room temperature, gently shaking during the reaction is preferred.

  4. Add equal volume of 100mM Tris-HCl Buffer (pH=7.5)+ 0.5% BSA.

  5. Store the mixture at 4oC, and it is ready to use. Note: If the target protein solution contains other proteins (such as BSA), purify the protein solution or it will interfere with the labeling reaction. Note: Small molecules such as sodium azide need to be removed. Note: Tris buffer and other amine compounds will also interfere with the labeling reaction.

Citations


View all 12 citations: Citation Explorer
Florfenicol binding to molecularly imprinted polymer nanoparticles in model and real samples
Authors: Caro, Nelson and Bruna, Tamara and Guerreiro, Antonio and Alvarez-Tejos, Paola and Garret{\'o}n, Virginia and Piletsky, Sergey and Gonz{\'a}lez-Casanova, Jorge and Rojas-G{\'o}mez, Diana and Ehrenfeld, Nicole
Journal: Nanomaterials (2020): 306
New method for immobilising diverse proteins onto cubic micro-protein polyhedrin crystals
Authors: Yuasa, Haruna and Kotani, Eiji and Mori, Hajime and Takaki, Keiko
Journal: Protein expression and purification (2019): 105531
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)
Spinal Cord Inflammation: Molecular Imaging after Thoracic Aortic Ischemia Reperfusion Injury
Authors: Albadawi, Hassan and Chen, John W and Oklu, Rahmi and Wu, Yue and Wojtkiewicz, Gregory and Pulli, Benjamin and Milner, John D and Cambria, Richard P and Watkins, Michael T
Journal: Radiology (2016): 152222
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

References


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Galactose oxidase action on galactose containing glycolipids--a fluorescence method
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Effects of chronic ethanol administration on the activities and relative synthetic rates of myelin and synaptosomal plasma membrane-associated sialidase in the rat brain
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Mitochondrial reactive oxygen species in mice lacking superoxide dismutase 2: attenuation via antioxidant treatment
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A new, sensitive method for enzyme kinetic studies of scarce glucosides
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Evaluation of assays for the measurement of bovine neutrophil reactive oxygen species
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Journal: Vet Immunol Immunopathol. (2006)
Kinetic analysis of semisynthetic peroxidase enzymes containing a covalent DNA-heme adduct as the cofactor
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K+-independent actions of diazoxide question the role of inner membrane KATP channels in mitochondrial cytoprotective signaling
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A Role for Mitochondrial Dysfunction in Perpetuating Radiation-Induced Genomic Instability
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