Preparing Cell Smears for Staining

Rodig, Scott J.

doi:10.1101/pdb.prot099630

Cited by 2 publications

(1 citation statement)

References 2 publications

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“…The cells were stained with Streptavidin-HRP and then incubated with TMB solution. The optical densities were read at 590 nm and their relative binding affinities were determined [24].…”

Section: Binding Affinityy Assaymentioning

confidence: 99%

Design, synthesis and analysis of charged RGD derivatives

Mujyambere

2023

Bioinformation

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In the present study, negatively charged N-Biotin-RGD and positively charged C-Biotin-RGD were designed, synthesized, and characterized with docking analysis. The fixation of MDA-MB-231 cells with formalin made their cell surface neutrally charged thus removing the electrostatic interactions between charged biotinylated RGD derivatives and MDA-MB-231 cells. The results of the binding affinity of biotinylated RGD derivatives against MDA-MB-231 cells showed that N-Biotin-RGD had higher binding affinity than C-Biotin-RGD. The cytotoxic effect was analyzed by incubating charged biotinylated RGD derivatives with live MDA-MB-231 cells. MDA-MB-231 cell surface is negatively charged due to high hypersialyliation of polyglycans and Warburg effect. The results of their cytotoxic activities against live MDA-MB-231 cells were found to be electrostatic in nature. C-Biotin-RGD had an attractive interaction with the MDA-MB-231 cell surface resulting in a higher cytotoxic effect. In comparison, N-Biotin-RGD had a repulsive interaction with the MDA-MB-231 cell surface resulting in a lower cytotoxic effect. Hence, positively charged C-Biotin-RGD is a better cytotoxic agent than a negatively charged N-Biotin-RGD against MDA-MB-231 cells.

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“…The cells were stained with Streptavidin-HRP and then incubated with TMB solution. The optical densities were read at 590 nm and their relative binding affinities were determined [24].…”

Section: Binding Affinityy Assaymentioning

confidence: 99%

Design, synthesis and analysis of charged RGD derivatives

Mujyambere

2023

Bioinformation

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Cell Staining

Rodig¹

2022

Cold Spring Harb Protoc

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When labeled antibodies are used to detect antigens in cells or tissues, several characteristics of an antigen can be readily determined. Most importantly, cell staining will show both the presence and subcellular localization of an antigen. Double-labeling techniques permit the simultaneous detection of two antigens, allowing comparisons of the relative distribution of different antigens. Many cell-staining methods can also be used in conjunction with conventional histological stains and autoradiographic methods to compare the localization of the antigen with other markers. Cell staining can also be used in pathology studies for determining such variables as the type of infectious organism, the progenitor of a neoplastic cell, or the presence of an inflammatory response. With certain modifications, the procedure can be used to purify cells on the basis of the antigenic composition of their cell surface. Cell staining is a versatile technique and, if the antigen is highly localized, can detect as few as a thousand antigen molecules in a cell or tissue. In some circumstances, cell staining may also be used to determine the approximate concentration of an antigen. Improvements in antibody labeling methods, microscopes, cameras, and image analyzers are rapidly extending the sensitivity of cell-staining procedures and are making these techniques more quantitative. Even without these improvements, cell staining can yield important qualitative and semiquantitative data. This introduction describes protocols for cell staining techniques and includes a discussion of major constraints, antibody selection, and troubleshooting.

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Preparing Cell Smears for Staining

Cited by 2 publications

References 2 publications

Design, synthesis and analysis of charged RGD derivatives

Design, synthesis and analysis of charged RGD derivatives

Cell Staining

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