Dead cells existing prior to the experiments were removed using Histopaque-1077 (Sigma) or Dead Cell Removal kit (Miltenyl Biotec). Freshly isolated intact iBCBL-1 cell lines were treated with 1 µg/ml Dox for the indicated times. Cell viability was then measured using trypan blue exclusion assays. For apoptosis assays, cells were analyzed by FITC-annexin V staining (BioLegend) or terminal deoxynucleotide transferase (TdT)-mediated dUTP nick labeling (TUNEL) assay kit (AAT Bioquest). All imaging and quantitation of apoptotic cells were performed using Cellometer Vision CBA Image Cytometry (Nexcelom), which employs four independent images for cell counting. FCS Express 6 Flow software (De Novo Software) was used for data analysis.
Tunel assay kit
Manufactured by AAT Bioquest
Sourced in United States
The TUNEL assay kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death, in cells. It works by labeling the fragmented DNA that is characteristic of apoptotic cells. The kit provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
Histopaque 1077, by Merck Group (1 mentions)
Fitc annexin 5 staining, by BioLegend (1 mentions)
Fcs express 6 flow software, by De Novo Software (1 mentions)
Dead cell removal kit, by Miltenyi Biotec (1 mentions)
Annexin 5 fitc apoptosis detection kit, by MBL Life Science (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Dmem high glucose, by GE Healthcare (1 mentions)
Fetal bovine serum (fbs), by Avantor (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions)
3 protocols using tunel assay kit
1
Apoptosis Quantification in iBCBL-1 Cells
2
Vitamin K2 Modulates Apoptosis in Cells
3
Inhibition of C3a Receptor Attenuates Endothelial Damage
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C3aR inhibitor (SB290157) was purchased from (Calbiochem, Darmstadt, Germany). Immortalized bEnd.3 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Dulbecco's Modified Eagle Medium (DMEM/High Glucose, GE, Utah, USA), Pen strep (Gibco, USA), FBS (VWR, USA) and all reagents used were AR and molecular grade. TUNEL assay kit was obtained from AAT Bioquest (USA). Antibodies (monoclonal anti-mouse Caspase3; monoclonal anti-mouse C3aR; monoclonal anti-mouse ICAM-1; monoclonal anti-mouse occludin; monoclonal anti-mouse Cox-2; monoclonal gp91-phox and MnSOD) were obtained from Santa Cruz Biotechnology, CA, USA. Recombinant C3a (rmC3a) was obtained from R&D system (USA). Lectin was purchased from Vector Lab (USA). Protease inhibitor cocktail was purchased from Sigma (MO). RIPA extraction buffer was procured from Fisher Scientific (USA). All secondary fluorescent antibodies were purchased from Invitrogen (USA).
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