BGC-823-miR-148a cells or BGC-vector cells were isolated and made into single cell suspensions, washed with PBS and then fixed in 3 ml cold 70% ethanol for 1 h at −20°C. The cells were then washed three times with BioLegend Cell Staining Buffer and resuspended in 100 μl of PBS. APC-conjugated Ki-67 antibody (10 μl, BioLegend) was added and incubated at room temperature in the dark for 30 min. Cells were washed with BioLegend Cell Staining Buffer and then resuspended in 0.5 ml of PBS for flow cytometric analysis.
Apc conjugated ki 67 antibody
Manufactured by BioLegend
The APC-conjugated Ki-67 antibody is a laboratory reagent used to detect the expression of the Ki-67 protein, a marker of cellular proliferation. This antibody is conjugated with the fluorescent dye Allophycocyanin (APC), allowing its detection by flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Cell staining buffer, by BioLegend (1 mentions)
Permeabilization buffer, by Thermo Fisher Scientific (1 mentions)
Percp cy5.5 conjugated cd4, by BioLegend (1 mentions)
Brilliant violet 510 conjugated cd8a, by BioLegend (1 mentions)
Fitc conjugated cd44, by BioLegend (1 mentions)
Pe conjugated cd62l, by BioLegend (1 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions)
2 protocols using apc conjugated ki 67 antibody
1
Cell Cycle Analysis by Flow Cytometry
2
Antigen-specific T cell proliferation assay
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Freshly isolated splenocytes (1 × 10E6) were seeded in 96-well plates and stimulated with 5 μg/mL Ag85B antigen or 1 μ/mL ESAT-6 antigen. As positive control, the cells were stimulated with 1 μg/mL α-CD3 (BioLegends). Antigen-specific T cell proliferation was analyzed after 6 days of incubation with antigens. The cells were blocked, as described above, and subsequently stained with PerCP/Cy5.5-conjugated CD4, Brilliant Violet 510™-conjugated CD8a, FITC-conjugated CD44, PE-conjugated CD62L and Brilliant Violet 421™- conjugated CD90.2 antibodies (all from BioLegends), all diluted 1:100. After staining, the cells were fixed using eBioscience™ Foxp3/Transcription Factor Staining Buffer Set and permeabilized using eBioscience™ Permeabilization Buffer, according to the manufacturer's instructions. Subsequently, cells were intracellularly stained with 1:50 diluted APC-conjugated Ki-67 antibody (BioLegends) and analyzed by flow cytometry.
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