NoW-Dtech™ – TR-FRET Assay Kit
Human IgM

Description

Accurate detection and quantification of human total IgM are important for a wide range of research applications, including biomarker studies, immunological research, and biopharmaceutical development. TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) provides a sensitive and homogeneous approach for quantitative protein detection based on the proximity of fluorescent donor and acceptor molecules. The NoW-Dtech™ Human IgM Kit enables the quantitative detection of human total IgM in serum, plasma, and cell culture supernatants using a fast, homogeneous, and wash-free mix-and-read assay format. The assay uses Bright-Dtech™ fluorescent lanthanide nanoparticles as long-lived fluorescent donors. Their time-resolved emission helps reduce short-lived background fluorescence, supporting sensitive and reliable detection in complex biological samples.

TR-FRET Assay Principle

The NoW-Dtech™ Human IgM assay uses two antibodies directed against human IgM. One antibody is coupled to Bright-Dtech™ lanthanide nanoparticles, which act as the fluorescent donor, while the second antibody is coupled to a fluorescent acceptor. When both antibodies bind to human IgM, the donor and acceptor are brought into close proximity, enabling fluorescence resonance energy transfer (FRET) from the donor to the acceptor. Following excitation, the resulting time-resolved fluorescence signal is measured using a compatible TR-FRET plate reader. The measured signal is proportional to the amount of human IgM present in the sample, enabling quantitative analysis.

 

Human IgM Kit: Protocol

The NoW-Dtech™ Human IgM Kit is designed for a simple mix-and-read workflow. After addition of the sample and detection reagents, the assay requires a 1.5 h incubation step, followed by fluorescence measurement. No washing or separation steps are required.

Please refer to the data sheet for detailed instructions and recommended assay conditions before use.

 

Human IgM Kit: Validation

Human IgM Kit: Required materials but not supplied

• Vials
• Precision single- and multichannel pipettes with disposable tips
• Adhesive sealing film or plate covers
• Plate reader with TR-FRET option

 

Human IgM Kit: Detection system

For optimal assay performance, fluorescence should be measured using a compatible time-resolved fluorescence plate reader with TR-FRET capability. Instrument settings may vary depending on the reader used.

Contact our team if you need assistance selecting a suitable detection system for your NoW-Dtech™ assay.

 

For all technical support, commercial enquiries or further information, contact the Poly-Dtech team.

 

 

Scientific Publications

Ultrabright Terbium Nanoparticles for FRET Biosensing and In Situ Imaging of Epidermal Growth Factor Receptors.
Design of a ligand that can be simultaneously applied as an efficient light-harvesting antenna for Tb surface ions and a strong binder of biomolecules to LnNPs surfaces.
Cyrille Charpentier, Vjona Cifliku, Joan Goetz, Aline Nonat, Clémence Cheignon, Marcelina Cardoso Dos Santos, Laura Francés-Soriano, Ka-Leung Wong, Loïc J Charbonnière, Niko Hildebrandt
Chemistry. 2020 Nov 17;26(64):14602-14611.
doi: 10.1002/chem.202002007

Live cell imaging without autofluorescence using terbium nanoparticles.
Application of a new type of surface photosensitized terbium NPs (Tb-NPs) for autofluorescence-free intracellular imaging in live HeLa cells. Combination of exceptionally high brightness, high photostability, and long photoluminescence (PL) lifetimes for highly efficient suppression of short-lived autofluorescence, enabling timed PL imaging of intracellular vesicles over 72 h without toxicity and at extremely low concentrations of Tb-NP for up to 12h.
Joan Goetz, Marcelina Cardoso Dos Santos, Ka-Leung Wong, Loïc J Charbonnière, Niko Hildebrandt, Hortense Bertenlian
Bioconjugate Chem. 2018, 29, 4, 1327–1334.
doi: 10.1021/acs.bioconjchem.8b00069