Lung cells were plated at 1 × 106 cells per well and stained with Fc block (CD16/32, 1:100, Tonbo Biosciences) for 30 min on ice. Cells were surface stained with the endothelial marker CD31 (1:100, clone: MEC3.1, eBiosciences), epithelial marker CD326 (1:100, clone: CD326, eBiosciences), and immune marker CD45 (1:100, clone: F11, eBiosciences) for 30 min on ice. The live/dead dye, SYTOX Blue (ThermoFisher), was added to cells and incubated for 3 min prior to sorting into 384-well plates (Bio-Rad Laboratories, Inc) prefilled with lysis buffer using the Sony LE-SH800 cell sorter (Sony Biotechnology Inc), a 100μm sorting chip (Catalog number: LE-C3110) and ultra-purity mode. Single color controls were used to perform fluorescence compensation and generate sorting gates. 384-well plates containing single cells were spun down, immediately placed on dry ice and stored at -80C.
Fc block cd16 32
Manufactured by Cytek Biosciences
The Fc block (CD16/32) from Cytek Biosciences is a laboratory equipment designed to block Fc receptors on cells, reducing nonspecific binding of antibodies. It serves as a tool for improving the specificity of immunoassays and flow cytometry experiments.
Automatically generated - may contain errors
3 protocols using fc block cd16 32
1
Single-cell sorting of lung cells
2
Single-cell sorting of lung cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Lungs were plated at 1 × 106 cells per well and stained with Fc block (CD16/32, 1:100, Tonbo Biosciences) for 30 min on ice. Cells were surface stained with the endothelial marker CD31 (1:100, eBioscience/ThermoFisher), epithelial marker Epcam (1:100, eBioscience/ThermoFisher), and immune marker CD45 (1:100, eBioscience/ThermoFisher) for 30 min on ice. The live/dead dye, Sytox Blue (Invitrogen), was added to cells and incubated for 3 min prior to sorting into 384-well plates (Bio-Rad Laboratories, Inc) prefilled with lysis buffer using the Sony LE-SH800 cell sorter (Sony Biotechnology Inc), a 100 μm sorting chip (Catalog number: LE-C3110) and ultra-purity mode. Single color controls were used to perform fluorescence compensation and generate sorting gates. 384-well plates containing single cells were spun down, immediately placed on dry ice and stored at − 80 ºC.
3
Single-cell sorting of lung cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lungs were plated at 1 × 106 cells per well and stained with Fc block (CD16/32, 1:100, Tonbo Biosciences) for 30 min on ice. Cells were surface stained with the endothelial marker CD31 (1:100, clone: MEC3.1, eBiosciences), epithelial marker Epcam (1:100, clone: CD326, eBiosciences), and immune marker CD45 (1:100, clone: F11, eBiosciences) for 30 min on ice. The live/dead dye, Sytox Blue (Invitrogen), was added to cells and incubated for 3 min prior to sorting into 384-well plates (Bio-Rad Laboratories, Inc) prefilled with lysis buffer using the Sony LE-SH800 cell sorter (Sony Biotechnology Inc), a 100 μm sorting chip (Catalog number: LE-C3110) and ultra-purity mode. Single color controls were used to perform fluorescence compensation and generate sorting gates. 384-well plates containing single cells were spun down, immediately placed on dry ice and stored at −80C.
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