TAMRA-PEG4-Alkyne

TAMRA-PEG4-Alkyne is a dye derivative of TAMRA (HY-135640) containing 4 PEG units. TAMRA-PEG4-Alkyne contains Alkyne groups and can undergo copper-catalyzed azide-alkyne cycloaddition (CuAAc) with molecules containing Azide groups (Ex/Em = 553/575 nm). In Vitro:Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Gel Fluorescence Analysis of Probe-Modified Proteins.
1.1 Incubate BSA or proteins extracted from MCF7 cells with sulfonated probe at 25 °C for 3 hours.
1.2 Add freshly premixed click chemistry reaction mixture (50 μM TAMRA-PEG4-Alkyne, 100 μM TBTA, 1 mM TCEP, and 1 mM CuSO4) to the above mixture.
1.3 Incubate at room temperature for 1 hour.
1.4 Then add 4× SDS-PAGE loading buffer and boil at 95°C for 10 minutes.
1.5 Analyze the samples using a 12% SDS-PAGE gel (polyacrylamide gel electrophoresis).

2. Cellular Imaging.
2.1 MCF7 cells were washed with PBS and pretreated with a sulfonium-based probe for 1 hour.
2.2 They were washed three times with PBS.
2.3 Then, they were fixed with 300 μL of 4% formaldehyde PBS solution for 15 minutes, followed by permeabilization with 300 μL of 0.1% Triton X-100 PBS solution for 15 minutes.
2.4 Freshly premixed click chemistry reaction mixture (10 μM TAMRA-PEG4-Alkyne (DMSO stock solution), 10 μM TBTA, 0.1 mM TCEP, and 0.1 mM CuSO4) was added to the mixture.
2.5 Incubation continued at room temperature for 1 hour.
2.6 The cells were washed three times with PBS, followed by one wash with 0.1% Tween PBS solution.
2.7 Finally, the cells were stained with 4′,6-diamidinyl-2-phenylindole (DAPI) for imaging.
2.8 The cells were observed using a confocal microscope or a fluorescence microscope.