Supernatant of hybridoma 2.4g2 (anti-CD16/32) was used to block Fc receptor. Cells were stained with Fixable Viability Dye eFluor 506 (eBioscience) to detect dead cells before surface staining. Annexin V apoptosis detection kit, eFlour 450 (eBioscience) and Ki-67 kit (BD Biosciences) were used according to manufacturers’ protocol. Antibodies used for flow cytometry were listed in Supplementary Table 1 . Pan-caspase activity was detected by FAM-FLICA™ Poly Caspase Assay Kit (ImmunoChemistry Technologies) according to manufacturer's protocol. For intracellular cytokine staining and transcription factor staining, Foxp3/Transcription Factor Staining Buffer Set (eBioscience) was used as manufacturer’s protocol. IELs (1 × 106 cells/ml) and LPLs (1 × 106 cells/ml) were stimulated with IL-12 (10 ng/mL) or IL-23 (10ng/mL)/IL-1β (10ng/mL) with GolgiPlug at 37°C for 4 hours for cytokine production, or IL-15 (20 ng/mL) at 37°C for 24 hours for Bcl-2 expression. Data were acquired on a FACSCanto II (BD Biosciences) and analyzed with FlowJo software (TreeStar).
Ki 67 kit
Manufactured by BD
The Ki-67 kit is a laboratory equipment product designed for the detection and quantification of the Ki-67 protein, which is a widely used marker for cell proliferation. The kit provides the necessary reagents and protocols to perform immunohistochemical or immunocytochemical analysis to determine the presence and distribution of Ki-67 in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2, by BD (3 mentions)
Eflour450, by Thermo Fisher Scientific (2 mentions)
Annexin 5 apoptosis detection kit, by Thermo Fisher Scientific (2 mentions)
Fixable viability dye efluor 506, by Thermo Fisher Scientific (2 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions)
Fam flica poly caspase assay kit, by ImmunoChemistry Technologies (2 mentions)
Live dead fixable red dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Fitc ki 67, by BD (1 mentions)
3 protocols using ki 67 kit
1
Multiparametric Flow Cytometry Analysis
2
LDL-induced HUVEC cell death and proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Confluent HUVECs were either treated or not treated with native LDL or Mox-LDL at a final concentration of 100 μg/mL for 24 hours. Cell death was detected using the LIVE/DEAD Fixable Red Dead Cell Stain Kit (Molecular Probes; Cat. number L-23102) following manufacturer's instructions. HUVECs were harvested and stained with the near-IR Dead Cell Stain which is excited at a 633/635 nm wavelengths. For proliferation analysis, BD Pharmingen Ki67 kit was used and the assay was also performed according to manufacturer's instructions. In brief, after staining with the Red Dead Cell Stain, cells were fixed, permeabilized, and then stained with FITC-Ki67 (BD Pharmingen, San Diego, CA, USA; Cat. number 612472) for 20 min at 4°C. Cells were then analyzed using BD FACSCanto II flow cytometry, and data were analyzed using FlowJO (Tree Star, inc.©). The experiments were performed in duplicate and the percentages of proliferating and dead cells were averaged between the duplicates.
3
Multiparametric Flow Cytometry Analysis
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