In solid white-bottom 96-well plates (#353296, Corning), 10,000 cells were plated and allowed to attach overnight. Cells were treated with vehicle, 10 μM enzalutamide, or 2 mM staurospaurine (positive control) in triplicate, and RealTime-Glo Annexin V Apoptosis and Necrosis kit (#JA1011, Promega; Madison, WI) was added per manufacturer instructions. Briefly, 50 μL containing 10,000 cells were added to each well of a 96-well plate. The next day, 50 μL of 4X concentrated drugs (vehicle, enzalutamide, or staurospaurine) and 100 μL of 2X detection reagents were added. Luminescence and fluorescence (485 nm/525 nm) were measured daily using the Synergy LX Multimode Reader (Biotek; Winooski, VT) and normalized to blank wells.
Realtime glo annexin 5 apoptosis and necrosis kit
Manufactured by Promega
Sourced in United States
The RealTime-Glo Annexin V Apoptosis and Necrosis kit is a laboratory reagent that can be used to detect and quantify cellular apoptosis and necrosis in real-time. The kit contains two cell-permeable reagents that bind to phosphatidylserine and enable the measurement of these processes in living cells.
Automatically generated - may contain errors
Lab products found in correlation
Solid white bottom 96 well plates, by Corning (2 mentions)
Synergy lx multi mode reader, by Agilent Technologies (2 mentions)
Azacitidine, by Selleck Chemicals (1 mentions)
Zebularine, by Selleck Chemicals (1 mentions)
Decitabine, by Selleck Chemicals (1 mentions)
Venetoclax, by Active Motif (1 mentions)
Triton x 100, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Texas red x phalloidin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta solution, by Merck Group (1 mentions)
Penicillin streptomycin solution, by Merck Group (1 mentions)
4 protocols using realtime glo annexin 5 apoptosis and necrosis kit
1
Apoptosis and Necrosis Assay Protocol
2
In vitro Sensitivity and Synergy in KMT2A-rearranged Infant ALL
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All in vitro sensitivity and synergy experiments were performed on an extensively characterized panel of eight patient-derived KMT2A-rearranged infant ALL cell lines (PER cell lines) [13 (link), 14 (link)], two commercially available KMT2A-rearranged infant ALL cell lines (ALL-PO and KOPN-8), and cells from four KMT2A-rearranged infant ALL patient-derived xenografts (MLL-5, MLL-7, MLL-14, and LR-iALL2) [15 (link), 16 (link)]. Assessment of in vitro sensitivity to azacitidine, decitabine, zebularine (Selleck Chemicals) and venetoclax (Active Biochem) and their synergy with each of the nine conventional chemotherapy agents used to treat infants with ALL, and synergy of each of the hypomethylating agents with venetoclax was performed as previously described [13 (link)]. In vitro drug combination studies were analyzed using SynergyFinder 2.0 [17 (link)]. Western blots were performed to detect DNMT1 levels following incubation with azacitidine and decitabine. Apoptosis and necrosis assays were performed using the RealTime-Glo™ Annexin V Apoptosis and Necrosis kit (Promega) according to the manufacturer’s instructions (Supplementary Methods ).
3
Apoptosis and Necrosis Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In solid white-bottom 96-well plates (#353296, Corning), 10,000 cells were plated and allowed to attach overnight. Cells were treated with vehicle, 10 μM enzalutamide, or 2 mM staurospaurine (positive control) in triplicate, and RealTime-Glo Annexin V Apoptosis and Necrosis kit (#JA1011, Promega; Madison, WI) was added per manufacturer instructions. Briefly, 50 μL containing 10,000 cells were added to each well of a 96-well plate. The next day, 50 μL of 4X concentrated drugs (vehicle, enzalutamide, or staurospaurine) and 100 μL of 2X detection reagents were added. Luminescence and fluorescence (485 nm/525 nm) were measured daily using the Synergy LX Multimode Reader (Biotek; Winooski, VT) and normalized to blank wells.
4
Cytotoxicity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Trypsin-EDTA solution, phosphate saline buffer (PBS), dimethyl sulfoxide (DMSO), fetal calf serum (FCS), penicillin/streptomycin solution, Triton X-100, DAPI, and MTT [3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide] reagent were purchased from Sigma Aldrich, Merck KgaA (Darmstadt, Germany). Dulbecco’s Minimum Essential Medium (DMEM; 30-2002™) was provided by ATCC (American Type Cell Collection, Lomianki, Poland). Texas Red™-X Phalloidin was provided by Thermo Fisher Scientific Inc. (Waltham, MA, USA). The RealTime-Glo™ Annexin V Apoptosis and Necrosis Kit was provided by Promega (Madison, WI, USA). Cytation 1 and Cytation 5 instruments, as well as the Gen5™ Microplate Data Collection and Analysis Software (Version 3.12), were bought from Biotek Instruments Inc. (Winooski, VT, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!