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

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

Red Fluorescence

Amplite® Fluorimetric Goat Anti-Rabbit IgG-HRP Conjugate ELISA Assay Kit *Red Fluorescence* is a fluorimetric assay kit for detection of rabbit IgG in ELISA assays using goat anti-rabbit 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 polyclonal antibody per well. AAT Bioquest
Workflow: Optimized for ELISA and compatible with HTS liquid handling.
Instrument Compatibility: Fluorescence detection at Ex/Em 540/590 nm; absorbance can also be read at 576 ± 5 nm.

Detection of rabbit total IgG using the Amplite® Fluorimetric ELISA Kit. Rabbit IgG was diluted into 1 µg/mL and made 1 to 3 serial dilutions in 0.2 M sodium bicarbonate buffer at pH 9.4. 100 µL/well serial dilutions were coated into a 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 using the reagents. 1 to 6000 dilutions of goat anti-rabbit IgG-HRP conjugate were used. The reactions were incubated for 15 to 60 minutes and then measured for fluorescence at Ex/Em = 540/590 nm with Gemini fluorescence microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
11541	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 Polyclonal antibody in the 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 the assays that detect goat anti-rabbit IgG as the secondary detection agent.

Citations

View all 10 citations: Citation Explorer

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

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

References

View all 61 references: Citation Explorer

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

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

Absorbance microplate reader
Absorbance	576 ± 5 nm
Recommended plate	Clear bottom

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

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Email	sales@aatbio.com
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Custom size	Inquire
Technical support	Contact us
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Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on October 13, 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-Rabbit IgG-HRP Conjugate	1 vial (25 µL)