ReadiUse™ Preactivated HRP NHS ester

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<p>Mechanism for conjugation of horseradish peroxidase (HRP) to target protein using activated NHS ester.</p>
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Unit Size: Cat No: Price (USD): Qty:
300 ug 11025 $95


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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

Storage Freeze (<-15 °C)
Category Enzyme Detection
Horseradish Peroxidase (HRP)
Related
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.




Protocol


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This protocol only provides a guideline, and should be modified according to your specific needs.
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.
Example data analysis and figures

Figure 1.

Mechanism for conjugation of horseradish peroxidase (HRP) to target protein using activated NHS ester.

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






Resources
 
Safety Data Sheet (SDS)


Documents
1. Enzyme Probes & Assay Kits

Certificate of Analysis