Description

The FLISA-Dtech™ Mouse TNF-Alpha (TNF-α) Kit enables the quantitative determination of TNF-α in mouse serum and plasma, supporting research applications involving inflammatory and immune responses.

Based on TR-FLISA (Time-Resolved Fluorescence Linked Immunosorbent Assay) technology, the assay combines the established ELISA format with the high signal intensity and long-lived fluorescence of Bright-Dtech™ fluorescent lanthanide nanoparticles. Time-resolved fluorescence (TRF) detection helps reduce short-lived background fluorescence, supporting sensitive and reliable quantitative detection.

The assay uses Bright-Dtech™ nanoparticles coupled to an anti-IgG antibody for fluorescence detection of the TNF-α-specific detection antibody, supporting sensitive detection of TNF-α in mouse serum and plasma.

TR-FLISA Assay Principle

The assay uses a sandwich FLISA format in which the microplate is pre-coated with an anti-TNF-α capture antibody. TNF-α present in the standards or diluted samples binds to the immobilized capture antibody.

ter incubation, an anti-TNF-α detection antibody is added and binds to the captured TNF-α, forming the sandwich complex. For detection, Bright-Dtech™ nanoparticles coupled to an anti-IgG antibody bind to the detection antibody, generating a fluorescent signal. The long-lived emission of the Bright-Dtech™ nanoparticles enables time-resolved fluorescence measurement using a compatible TRF reader.

The fluorescence signal is proportional to the amount of TNF-α captured in the assay, allowing its concentration to be determined using a standard curve.

Mouse TNF-α Kit: Protocol

The FLISA-Dtech™ Mouse TNF-α Kit uses a simple plate-based workflow. The pre-coated and blocked microplate is first prepared according to the protocol, followed by addition of standards or diluted samples. After incubation, the anti-TNF-α secondary antibody is added, followed by Bright-Dtech™–anti-IgG detection reagent. Following the final incubation and washing steps, the fluorescence signal is measured using a compatible TRF reader.

 

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

Mouse TNF-α Kit: Validation

 

Mouse TNF-α Kit: Required materials

• Deionized or distilled water
• Precision single- and multichannel pipettes with disposable tips
• Adhesive sealing film or plate covers
• Automatic plate washer
• Plate reader with TRF option

 

Mouse TNF-α Kit: Detection system

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

Contact our team if you need assistance selecting a suitable detection system for your FLISA-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