Conjugated SNAP-Antibody Suitable for Live Cell Imaging

June 15, 2023

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Quality of Results

Ease-of-Optimization

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Company:

New England Biolabs

Product Name:

SNAP-Cell® 647-SiR

Catalog Number:

S9102S

Clone Number:

N/A

This research involves studying how D1 dopamine receptor, a member of the GPCR superfamily, can signal from distinct membrane compartments within the cell. We have found that D1 dopamine receptor can be activated at the plasma membrane, endosomes, and Golgi membrane. SNAP-Cell-647, which is membrane permeant, was used in cells expressing a SNAP-tagged D1 dopamine receptor on the cell surface and at internal membrane compartments to visualize receptor via live-cell fluorescence confocal microscopy.

Experimental Design and Results Summary

Applications

Live-cell fluorescence microscopy

Sample

HeLa cell

Primary Incubation

1:3000, 37degrees C, 30 min

Blocking Agent

Serum-free media (DMEM or DMEM:F12 no phenol red)

Secondary Incubation

Serum-free media 3x wash, 37 C, 30 min incubation in 3rd wash

Tertiary Incubation

N/A

Detection

Fluorescence microscope

Results Summary

This product helped us detect SNAP-labelled dopamine receptor in live-cells that we could image on a confocal fluorescence microscope. Since this dye is membrane impermeant, we were able to identify receptor at both the plasma membrane and internal compartments within cells. Important to have a long enough incubation in dye, then wash 3 times and incubate in the 3rd wash for 20-30 minutes at 37 degrees.

DOI or PMID #

N/A

Additional Notes

Can do shorter incubations

Image Gallery

Summary

The Good

Very strong binding of conjugated dye to SNAP tag; can tag internal proteins in live cells

The Bad

takes some optimization; sometimes is too internalized and does not properly label cell-surface SNAP-tagged protein

The Bottom Line

Great product for imaging internally-localized SNAP-labelled proteins, must be diligent with washes and incubations in order to get proper labelling.

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