The Twinkle Factory reagents

^match540

ref: match540-50X

a fluorogenic dimerizer emitting at 540 nm
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^match550

ref: match550-50X

the most powerful of our fluorogenic dimerizers, emitting at 550 nm

^match600

ref: match600-50X

a fluorogenic dimerizer emitting at 600 nm
–

Cognate proteic tags

All above molecular glues have been optimized for ^FIREmate and ^FIREtag

^TFLime

ref: 480541-250

A greenish, membrane-permeant, fluorogenic ligand

a.k.a. HMBR

Absorption and emission wavelengths when bound to
FAST1: 481 / 540 nm – FAST2: 480 / 541 nm – pFAST : 484 / 550 nm

Molar absorption coefficient when bound to
FAST1: 45,000 – FAST2: 41,000 – pFAST: 56,000 M–1cm–1

Fluorescence quantum yield when bound to
FAST1: 23 % – FAST2: 23 % – pFAST: 13%

Affinity constant at 25°C of its complex with
FAST1: 0.13 µM – FAST2: 0.07 µM – pFAST: 0.50 µM

^TFAmber

ref: 499558-250

A yellowish, membrane-permeant, fluorogenic ligand

a.k.a. HBR-3,5DM

Absorption and emission wavelengths when bound to
FAST1: 499  / 562 nm – FAST2: 499 / 558 nm

Molar absorption coefficient when bound to
FAST1: 48,000 – FAST2: 46,000 – pFAST: 55,000 M–1cm–1

Fluorescence quantum yield when bound to
FAST1: 49 % – FAST2: 57 % – pFAST: 50%

Affinity constant at 25°C of its complex with
FAST1: 0.08 µM – FAST2: 0.06 µM – pFAST: 0.80 µM

^TFCoral

ref: 516600-250

A red, membrane-permeant, fluorogenic ligand

a.k.a. HBR-3,5DOM

Absorption and emission wavelengths when bound to
FAST1: 518  / 600 nm – FAST2: 516 / 600 nm – pFAST: 525 / 600 nm

Molar absorption coefficient when bound to
FAST1: 39,000 – FAST2: 38,000 – pFAST: 51,000 M–1cm–1

Fluorescence quantum yield when bound to
FAST1: 31 % – FAST2: 40 % – pFAST: 19%

Affinity constant at 25°C of its complex with
FAST1: 0.97 µM – FAST2: 0.41 µM – pFAST: 3.9 µM

^TFAmber-NP

ref: 505559-250

A yellowish, non-membrane permeant, fluorogenic ligand

a.k.a. HBRAA-3E

Absorption and emission wavelengths when bound to
FAST1: 505  / 559 nm

Molar absorption coefficient when bound to
FAST1: 61,000 M–1cm–1

Fluorescence quantum yield when bound to
FAST1: 8 %

Affinity constant at 25°C of its complex with
FAST1: 1.3 µM

^TFFox-NP

ref: 524578-250

A red, non-membrane permeant, fluorogenic ligand

a.k.a. HBR-3,5DM

Absorption and emission wavelengths when bound to
pFAST: 524  / 578 nm

Molar absorption coefficient when bound to
pFAST: 58,000 M–1cm–1

Fluorescence quantum yield when bound to
pFAST: 22 %

Affinity constant at 25°C of its complex with
pFAST: 1.8 µM

^TFDarth

ref: 5140000-250

A dark, non-fluorogenic, ligand (a fluorescence quencher)

a.k.a. HBIR-3M

Affinity constant at 25°C of its complex with
pFAST: 0.005 µM

Cognate proteic tags

A wide variety of proteic tags have been developed during the recent years, either improving the properties of basic tags or featuring special properties.  As a result, we today recommend pFAST (brighter and more promiscuous than FAST1 or FAST2) for straightforward protein reporting or splitFAST2 (brighter and lower affinity than splitFAST) for protein-protein interaction reporting.  Most of our Fluorogens can be used with any version of FAST or splitFAST.  Besides, greenFAST is specific for ^TFLime and redFAST for ^TFCoral.  Used together, they hence allow multicolor orthogonal labeling.  frFAST was specifically designed with ^TFPoppy for far-red labeling.

The herebelow table provides guidance and references for helping your choice.