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ReadiUse™ Preactivated HRP NHS ester

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

References

View all 61 references: Citation Explorer
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)
Real-time assessment of spatial and temporal coupled catalysis within polyelectrolyte microcapsules containing coimmobilized glucose oxidase and peroxidase
Authors: Stein EW, Volodkin DV, McShane MJ, Sukhorukov GB.
Journal: Biomacromolecules (2006): 710
O2 Delivery and Redox State are Determinants of Compartment-specific Reactive Oxygen Species in Myocardial Reperfusion
Authors: Stoner JD, Clanton TL, Aune SE, Angelos MG.
Journal: Am J Physiol Heart Circ Physiol. (2006)
Fibrillar beta-amyloid peptide Abeta1-40 activates microglial proliferation via stimulating TNF-alpha release and H2O2 derived from NADPH oxidase: a cell culture study
Authors: Jekabsone A, M and er PK, Tickler A, Sharpe M, Brown GC.
Journal: J Neuroinflammation (2006): 24
Biochemical activity of reactive oxygen species scavengers do not predict retinal ganglion cell survival
Authors: Schlieve CR, Lieven CJ, Levin LA.
Journal: Invest Ophthalmol Vis Sci (2006): 3878
Page updated on November 23, 2024

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Catalog Number11025
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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

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12171501