JF635-HTL
JF635-HTL is a Fluorescent dye for spatiotemporally controlled live cell imaging and single-molecule localization microscopy SMLM. Its detection mechanism depends on the conformational change of a photoswitchable HaloTag psHaloTag, which integrates the light-responsive AsLOV2 domain: in the dark state, the folded Jα helix of AsLOV2 maintains the dye in a predominantly closed, non-fluorescent form; upon 450 nm illumination, a metastable photo-adduct forms between a cysteine side chain and the FMN cofactor of AsLOV2, causing undocking and unfolding of the Jα helix, which propagates a conformational change to the HaloTag near the dye binding site, shifting the dye's equilibrium to the open, fluorescent form; this process is fully reversible in the dark as the Jα helix refolds spontaneously, returning the dye to the closed, non-fluorescent state. For psHaloTag1a labeled with JF635-HTL, the excitation/emission wavelengths for the ON state are Ex/Em = 642/655 nm, while for psHaloTag1b labeled with JF635-HTL, the wavelengths are Ex/Em = 639/655 nm; when bound to wild-type HaloTag, the wavelengths are Ex/Em = 640/656 nm.
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- CAS. Nr.: 1811541-14-6
- Formel: C39H46ClF2N3O5Si
- Molecular Weight:738.33
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Speicherung:
Please store the product under the recommended conditions in the Certificate of Analysis.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biologische Aktivität
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Chemical Information
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Biologische Aktivität
Beschreibung
In Vitro
Guide
(The following is our recommended protocol. This protocol is for guidance only and should be modified according to your specific needs).
1. Stock Solution Preparation
(No stock solution details provided in literature, so this section is omitted)
2. Working Solution Preparation
2.1 Working concentration: 1 μM
2.2 Note: Adjust working solution concentration as needed; prepare fresh before use.
3. Staining Procedure
3.1 Sample type: Adherent cells (U2OS cells expressing psHaloTag1a targeted to nucleus, mitochondria, or F-actin)[1]
3.2 Incubation conditions: Incubate with 1 μM JF635-HTL for 2 h
3.3 Washing steps: Wash cells after incubation to remove unbound dye
4. Controls
4.1 Set up positive control: HaloTag-expressing cells labeled with JF635-HTL
5. Detection & Analysis
5.1 Instrument: Widefield or confocal fluorescence microscope
5.1.1 Ex/Em wavelength: 450 nm for photoactivation
5.2 Result analysis:
5.2.1 Fluorescence intensity changes: Deep-red fluorescence increases 3.5–4.1-fold after 450 nm illumination; fluorescence reverses to basal level in dark
5.2.2 Fluorescence localization: Fluorescence colocalizes with target subcellular structures (nucleus, mitochondria, F-actin)
5.2.3 Color changes: Deep-red fluorescence in the ON state after 450 nm illumination
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS. Nr. 1811541-14-6
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Molecular Weight 738.33
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Formel C39H46ClF2N3O5Si
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SMILES
O=C1OC2(C3=CC(C(NCCOCCOCCCCCCCl)=O)=CC=C13)C4=CC=C(N5CC(C5)F)C=C4[Si](C)(C)C6=CC(N7CC(C7)F)=CC=C62
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Versand
Room temperature in continental US; may vary elsewhere.
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Speicherung
Please store the product under the recommended conditions in the Certificate of Analysis.
Reinheit & Dokumentation
Verweise
Calculators
Konzentration (Stammlösung) × Volumen (Stammlösung) = Konzentration (Ziellösung) × Volumen (Ziellösung)
JF635-HTL Related Classifications
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