Amplite® Rapid Colorimetric APC-Maleimide Quantitation Kit

Amplite® Rapid Colorimetric APC-Maleimide Quantitation Kit is a colorimetric assay kit for quantification of maleimide groups on maleimide-activated APC proteins.

Detection Principle: Determines the maleimide-to-APC ratio from absorbance at 555 nm and 650 nm.
Workflow: Removes unreacted sensor with a spin column before absorbance measurement.
Instrument Compatibility: Compatible with cuvettes, NanoDrop™ spectrophotometers, and 96-well absorbance plate readers.
Applications: Supports rapid quantification of maleimide modification on APC proteins for conjugate characterization.

The Amplite® Rapid Colorimetric APC-Maleimide Quantitation Assay Principle.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
5527	2 Tests	Price

Overview

Maleimide-based crosslinking reagents are commonly used for conjugating proteins to other proteins or biomolecules. A significant challenge in maleimide chemistry is the precise quantification of maleimide moieties conjugated to proteins. The Amplite® Rapid Colorimetric APC-Maleimide Quantitation Kit facilitates the quantification of maleimide groups on maleimide-activated APC proteins using a proprietary sensor, Maleimide 555™, which has an absorption peak at ~555 nm. The assay involves the reaction of Maleimide 555™ with the maleimide-activated APC, followed by the separation of the reaction mixture via a spin column to remove the unreacted sensor. Subsequently, the absorption spectrum of the isolated product is measured and the amount of maleimide to APC is determined from the absorbance ratios at 555 nm and 650 nm, with the latter being the peak absorbance of APC. This quantitation kit is compatible with various detection systems, including traditional cuvettes, NanoDrop™ Spectrophotometers, and 96-well absorbance plate readers. It offers a robust and adaptable approach for the rapid quantification of maleimide modifications on APC proteins, addressing a critical need in protein biochemistry.

References

View all 50 references: Citation Explorer

Response to the Comment on "DpCoA tagSeq: Barcoding dpCoA-Capped RNA for Direct Nanopore Sequencing via Maleimide-Thiol Reaction".

Authors:

Shao, Xiaojian and Zhang, Hailei and Zhu, Zhou and Ji, Fenfen and He, Zhao and Yang, Zhu and Xia, Yiji and Cai, Zongwei

Journal:

Analytical chemistry (2024): 610-613

Maleimide-Thiol Linkages Alter the Biodistribution of SN38 Therapeutic Microbubbles Compared to Biotin-Avidin While Preserving Parity in Tumoral Drug Delivery.

Authors:

Ingram, Nicola and Abou-Saleh, Radwa H and Race, Amanda D and Loadman, Paul M and Bushby, Richard J and Evans, Stephen D and Coletta, P Louise

Journal:

Pharmaceutics (2024)

Identification of Low-Density Lipoprotein Receptor-Related Protein 1 as a CXCL14 Receptor Using Chemically Synthesized Tetrafunctional Probes.

Authors:

Miyajima, Rin and Tanegashima, Kosuke and Naruse, Naoto and Denda, Masaya and Hara, Takahiko and Otaka, Akira

Journal:

ACS chemical biology (2024): 551-562

Comment on "DpCoA tagSeq: Barcoding dpCoA-Capped RNA for Direct Nanopore Sequencing via Maleimide-Thiol Reaction".

Authors:

Vinther, Jeppe

Journal:

Analytical chemistry (2024): 606-609

Surface passivation and functionalisation for mass photometry.

Authors:

Sülzle, Jenny and Elfeky, Laila and Manley, Suliana

Journal:

Journal of microscopy (2024)

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	900 nm to 250 nm
Recommended plate	Clear bottom

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Page updated on October 25, 2026

Component A: Maleimide™ 555	2 Vials (1 vial is for 100 µg of APC-maleimide)
Component B: Assay Buffer	1 Bottle (15 mL)
Component C: Spin Column	2 Columns