Cell surface labelling

Figure 3. E. chrysanthemi cell surface labelling with sulfo-NHS-biotin. After labelling, the proteins were separated by SDS-PAGE, blotted onto nitrocellulose and revealed with PemB-antibodies (A) or with streptavidin-peroxidasc (B). Lane 1 A350 (wild type) lane 2 A837 kdgRy, lane 3 A350/pPME6. An arrowhead indicates the PemB position.

Yu J, Choi S, Richards Cl, Antoku Y, Dickson RM (2008) Live cell surface labeling with fluorescent Ag nanocluster conjugates. Photochem Photobiol 84 1435-1439... 

Tolsen, N. D., Boothroyd, B., and Hopkins, C. R. (1981) Cell-surface labelling with gold colloidal particles, the use of avidin and staphylococcal protein A-coated gold in conjunction with biotin and Fc-beanng ligands. J. Microsc 123,215—226. 

When HeLa cells were.cultured in medium supplemented with 5 mM sodium butyrate, their content of GM3 increased (Fig.2a). Increases varied from 3.5 to 5-fold depending on the experiment (4,8,12,13). When the butyrate was removed and the cells were cultured in normal medium for 24 h, the GM3 levels returned to those found in untreated cells (Fig. 2a). Similar results were observed when N-[acetyl-3H]-D-mannosamine, a precursor of sialic acid, was also included in the culture medium. In the butyrate-treated cells, radioactivity associated with GM3 increased 6.5-fold and 24 h after butyrate was removed, the amount of labeling returned to control values (Fig. 2b). We also were able to label the GM3 by means of a cell surface labeling technique. Control and butyrate-treated cells were exposed to 10 mM sodium periodate and the oxidized sialyl residues were reduced with NaBSfy. There was 5.5-fold more 3h associated with the GM3 recovered from the butyrate-... 

The following procedure has been routinely used in our laboratories to prepare immunolatex conjugates of high immunochemical activity for applications in cell surface labeling. 

Link AJ, Tirrell DA. Cell surface labeling of Escherichia coli via copper(I)-catalyzed [3 4- 2] cycloaddition. J. Am. Chem. Soc. 2003 125 11164-11165. 

Reolon LA, Martello CL, Schrank IS, Ferreira HB. Survey of surface proteins from the pathogenic Mycoplasma hyopneumoniae strain 7448 using a biotin cell surface labeling approach. Plos One. 2014 9(ll) ell2596. doi 10.1371/journal.pone.0112596. 

Labelling of antibodies with tetramethylrhodamine-5-isothicKyanate (TRITC) 357 Cell surface labelling 359... 

Further studies using NeuSAc have been related to the metabolism of extracellular CMP-NeuSAc and the detection of ectosialyltransferase activity. On the basis of a lag period observed for NeuSAc uptake and incorporation into glycoconjugates, and the absence of such a lag for CMP-NeuSAc incorporation, two routes have been proposed. The uptake of NeuSAc and metabolism as described above has been studied in hamster and mouse fibroblasts, and cell surface labelling of glycoprotein and glycolipid demonstrated (Datta 1974). The breakdown of CMP-NeuSAc was shown, and incorporation due to NeuSAc uptake rather than direct CMP-NeuSAc transfer proposed (Hirschberg et al. 1976). The uptake of CMP-NeuSAc into the cells (NIL, BHK and 3T3 fibroblasts) could be ruled out, and the K , for NeuSAc uptake was estimated to be 10 mM. Other experiments with CMP-NeuSAc and intact cell cultures (Painter and White 1976, Cerven 1977, see section III.9) pointed to surface sialyltransferase. Further studies by Fan and Datta (1980) provided evidence that both transfer and transport occur, by localization of acceptors within the cell and on the cell surface (plasma membrane), and direct demonstration of the presence of a plasma membrane sialyltransferase. The sialylation due to NeuSAc uptake occurs (at least initially) with different acceptors in comparison with CMP-NeuSAc plasma membrane sialylation. 

Peptide and protein labeling, cyclization and immobilization, protein-protein fusion, cell surface labeling [32,33]... 

Key words Single particle tracking. Diffusion coefficient. Confinement, Dwell time. Quantum dot. Live cell. Surface labeling. Videomicroscopy, Synapse... 

Figure 6.14 Amphiphilic complexes as luminescent cell surface labels. Reprinted with permission from [53]. Copyright 2011, American Chemical Society...

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