Amplite® Fluorimetric Goat Anti-Mouse IgG-HRP Conjugate ELISA Assay Kit *Red Fluorescence*

Amplite® Fluorimetric Goat Anti-Mouse IgG-HRP Conjugate ELISA Assay Kit

Red Fluorescence

Amplite® Fluorimetric Goat Anti-Mouse IgG-HRP Conjugate ELISA Assay Kit is a fluorimetric assay kit for detection of mouse IgG in ELISA assays using goat anti-mouse IgG-HRP as the secondary detection reagent.

Detection Principle: Detects HRP activity using a fluorogenic peroxidase substrate.
Sensitivity: Detects as little as 10 pg of monoclonal antibody per well.
Workflow: Optimized for ELISA and compatible with HTS liquid handling.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm with a microplate reader.

Detection of mouse total IgG using the Amplite® Fluorimetric Goat Anti-Mouse IgG-HRP ELISA Kit. Mouse IgG was diluted into 3 µg/mL and made 1 to 3 serial dilutions in 0.2 M sodium bicarbonate buffer, pH 9.4. 100 µL/well serial dilutions were coated into a solid black 96-well plate at 4 °C overnight, and blocked with 3% milk in PBS and 0.02% Tween-20 at 4 °C overnight. The wells were washed and assayed by using the reagents. 1 to 5000 dilutions of goat anti-mouse IgG-HRP conjugate were used. The reactions were incubated for 10 to 60 minutes and then measured for fluorescence at Ex/Em = 540/590 nm using Gemini fluorescence microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
11540	1000 Tests	Price

Overview

Horseradish Peroxidase (HRP) is a small molecule (MW ~40 KD) that is widely used in a variety of biological detections. HRP conjugates are extensively used as secondary detection reagents in ELISAs, immuno-histochemical techniques, Northern, Southern and Western blot analyses. Due to its small size, it rarely causes steric hindrance problem with antibody/antigen complex formation. It is usually conjugated to an antibody in a 4:1 ratio. Additionally, HRP is inexpensive compared to other labeling enzymes. The major disadvantage associated with peroxidase is their low tolerance to many preservatives such as sodium azide that inactivates peroxidase activity even at low concentration. Our Amplite® Fluorimetric ELISA Assay Kit contains all the essential components including our fluorogenic Amplite® Red HRP substrate for ELISA detection. The kit provides an optimized assay protocol that is compatible with HTS liquid handling, as little as 10 pg of a monoclonal antibody/well of a microplate can be detected. Its signal can be easily read by either fluorescence microplate reader or absorbance microplate reader. It can be used for assays that detect goat anti-mouse IgG as the secondary detection agent.

Citations

View all 11 citations: Citation Explorer

The combination of circRNA-UMAD1 and Galectin-3 in peripheral circulation is a co-biomarker for predicting lymph node metastasis of thyroid carcinoma

Authors:

Yu, Wenbin and Ma, Bo and Zhao, Wei and Liu, Jingtao and Yu, Hao and Tian, Zhihua and Fan, Zhiqin and Han, Haibo

Journal:

American Journal of Translational Research (2020): 5399

Assessment of Tofacitinib and Ruxolitinib and their Anti Inflammatory Effects on Myeloperoxidase

Authors:

Milton, Amber

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

References

View all 61 references: Citation Explorer

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

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

Spectral properties

Excitation (nm)	571
Emission (nm)	584

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	540 nm
Emission	590 nm
Cutoff	570 nm
Recommended plate	Solid black

Telephone
Fax
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Technical support	Contact us
Request quotation	Request
Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on October 26, 2026

Component A: Amplite® Red Peroxidase Substrate	2 vials
Component B: H2O2	1 vial (3% stabilized solution, 500 µL)
Component C: Assay Buffer	1 bottle (100 mL)
Component D: DMSO	1 vial (1 mL)
Component E: Goat Anti-Mouse IgG-HRP Conjugate	1 vial (25 µL)