Cells were harvested using non-enzymatic dissociation solution (Corning) and washed once with PBS. For staining of ICAM-1, cells were incubated in Human Fc Block (BD) for 10 min. Thereafter, cells were stained with FITC-conjugated antibody to ICAM-1 in the presence of Fc Block for 30 min at 4 °C. Cells were then washed 3× with PBS and analyzed by FACS Caliber (BD). Data analysis was performed using Flowjo V10 software (Ashland).
Non enzymatic dissociation solution
Manufactured by Corning
Non-enzymatic dissociation solution is a laboratory reagent used to facilitate the separation and dissociation of cells in cell culture applications. It is a chemically-based solution designed to disrupt cell-to-cell and cell-to-surface adhesions without the use of enzymes. The core function of this product is to provide a gentle and efficient method for harvesting and passaging cultured cells.
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Lab products found in correlation
3 protocols using non enzymatic dissociation solution
1
Quantifying Cell Surface ICAM-1 Expression
2
Quantifying Cell Surface ICAM-1 Expression
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Cells were harvested using non-enzymatic dissociation solution (Corning) and washed once with PBS. For staining of ICAM-1, cells were incubated in Human Fc Block (BD) for 10 min. Thereafter, cells were stained with FITC-conjugated antibody to ICAM-1 in the presence of Fc Block for 30 min at 4 °C. Cells were then washed 3× with PBS and analyzed by FACS Caliber (BD). Data analysis was performed using Flowjo V10 software (Ashland).
3
Immune Cell Phenotyping by Flow Cytometry
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Cells were harvested using non-enzymatic dissociation solution (Corning, Corning, NY) and washed once with PBS. For intracellular staining of CD68, cells were treated using the Fixation/Permeabilization Solution Kit (BD Biosciences, San Jose, CA), while all other surface stains proceeded to the following step. Cells were then incubated in Human Fc Block (BD) for 10 min. Thereafter, cells were stained with FITC, PE, or APC-conjugated antibody to CD68, CD80, CD86, HLA-ABC, HLA-DR, CCR5, CD163, or CD206 in the presence of Fc Block for 30 min at 4 °C. Cells were then washed with 3× with PBS and analyzed by flow cytometry.
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