Amplite® Human Serum Albumin (HSA) Site II Binding Assay Kit

Amplite® Human Serum Albumin (HSA) Site II Binding Assay Kit is a fluorimetric assay kit for measuring small-molecule binding to Site II of human serum albumin in a high-throughput microplate format.

Detection Principle: Measures displacement of HSA Blue™ S2 from HSA Site II by competing small molecules.
Signal Advantage: HSA Blue™ S2 shows a large fluorescence difference between HSA-bound and unbound states.
Workflow: Incubates for 15–45 minutes at room temperature and is designed for high-throughput screening.
Instrument Compatibility: Fluorescence detection at Ex/Em 365/480 nm with a 435 nm cutoff.

Response of Warfarin (Site-1 drug) and Ibuprofen (Site-2 drug) was measured using Amplite® Human Serum Albumin (HSA) Site II Binding Assay Kit. The response was acquired using Spectra Max Gemini XS (Molecular devices) with Ex/Em = 365/480 nm with cutoff = 435 nm.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
25402	200 Tests	Price

Overview

Human serum albumin (HSA) is one of the most important carriers for acidic drugs in human plasma and has been shown to bind a large number of different compounds in a reversible manner. Several different ligand binding sites have been identified for HSA. Among them, Site II has been identified as one of major drug binding sites. Amplite®™ Human Serum Albumin (HSA) Site II Binding Assay Kit is a fluorescence-based high throughput assay to determine the small molecule binding towards HSA. This assay is based on a novel fluorescent probe, HSA Blue™ S2. It has been characterized to bind to the site 2 of HSA with unique spectroscopic and binding properties. HSA Blue™ S2 displays a large fluorescence intensity difference between the protein-bound and protein-unbound state. The competition of small molecules for HSA binding in the presence of HSA Blue™ S2 results in low fluorescence intensities. This assay can be used as a high throughput screen tool to determine total binding to HSA at Site II.

References

View all 50 references: Citation Explorer

Anionic versus neutral Pt(II) complexes: The relevance of the charge for human serum albumin binding.

Authors:

Ricciardi, Loredana and Guzzi, Rita and Rizzuti, Bruno and Ionescu, Andreea and Aiello, Iolinda and Ghedini, Mauro and La Deda, Massimo

Journal:

Journal of inorganic biochemistry (2020): 111024

Human Serum Albumin Binding in a Vial: A Novel UV-pH Titration Method To Assist Drug Design.

Authors:

Dargó, Gergő and Bajusz, Dávid and Simon, Kristóf and Müller, Judit and Balogh, György T

Journal:

Journal of medicinal chemistry (2020): 1763-1774

Anticancer activity, calf thymus DNA and human serum albumin binding properties of Farnesiferol C from Ferula pseudalliacea.

Authors:

Tanzadehpanah, Hamid and Mahaki, Hanie and Samadi, Pouria and Karimi, Jamshid and Moghadam, Neda Hosseinpour and Salehzadeh, Sadegh and Dastan, Dara and Saidijam, Massoud

Journal:

Journal of biomolecular structure & dynamics (2019): 2789-2800

An integrated quantitative structure and mechanism of action-activity relationship model of human serum albumin binding.

Authors:

Serra, Angela and Önlü, Serli and Coretto, Pietro and Greco, Dario

Journal:

Journal of cheminformatics (2019): 38

The displacement study of 99m Tc-DTPA-Human serum albumin binding in presence of furosemide and metformin by using equilibrium dialysis and molecular docking.

Authors:

Chemlal, Laila and Akachar, Jihane and Makram, Sanaa and Zoubir, Brahim and Cherrah, Yahia and Eljaoudi, Rachid and Ibrahimi, Azeddine and Faouzi, Mly A

Journal:

IUBMB life (2019): 2003-2009

Spectral properties

Excitation (nm)	337
Emission (nm)	495

Usage and storage

Intended use

Research Use Only (RUO)

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Page updated on October 18, 2026

Component A: HSA Blue™ S2	1 vial
Component B: HSA Assay Buffer	1 vial (30 mL)
Component C: Human Serum Albumin solution (HSA)	1 vial (500 μL)
Component D: DMSO	1 vial (100 μL)