estradiol (E2) (Sigma-Aldrich, St. Louis, MO); 4-hydroxytamoxifen (4OHT) (Sigma-Aldrich); ICI 182,780 (fulvestrant) (Tocris Bioscience, Bristol, UK); staurosporine (STS) (Tocris Bioscience); and Z-VAD (Tocris Bioscience). E2, 4OHT, and fulvestrant were solubilized in 200-proof ethanol prior to use. STS and Z-VAD were solubilized in sterile DMSO.
Z vad
Manufactured by Bio-Techne
Sourced in United Kingdom
Z-VAD is a broad-spectrum caspase inhibitor that can be used to inhibit apoptosis in various cell types. It is a potent, cell-permeable, irreversible inhibitor of caspases.
Automatically generated - may contain errors
Lab products found in correlation
Estradiol e2, by Merck Group (1 mentions)
Ici 182 780 fulvestrant, by Bio-Techne (1 mentions)
4 hydroxytamoxifen 4 oht, by Merck Group (1 mentions)
Cell fractionation kit, by Cell Signaling Technology (1 mentions)
Azd2461, by Selleck Chemicals (1 mentions)
Vx 765, by Selleck Chemicals (1 mentions)
Rapamycin, by Merck Group (1 mentions)
Digitonin, by Merck Group (1 mentions)
Trehalose, by Merck Group (1 mentions)
Protein a sepharose, by Roche (1 mentions)
Ly294002, by Bio-Techne (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Apc anti human cd124 il 4rα, by BioLegend (1 mentions)
Igg isotype antibody, by BioLegend (1 mentions)
Glosensor, by Promega (1 mentions)
D luciferin, by PerkinElmer (1 mentions)
Annexin 5 pi apoptosis detection kit, by BD (1 mentions)
5 protocols using z vad
1
Estrogen Receptor Signaling Assay
2
ARPE-19 and Primary Human RPE Cell Fractionation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ARPE-19 cells were grown and maintained in T-75 mM flasks in DMEM F-12 medium containing 10% FBS and incubated at 37 °C with a constant supply of 5% CO2. Similarly, primary human RPE cells were grown in EMEM medium containing 10% FBS and 5% fetal calf serum at 37 °C and 5% CO2. Cytoplasmic and nuclear fractions were separated by using the Cell Fractionation Kit of Cell Signaling Technology (Danvers, MA, USA) following the protocol supplied by the manufacturers. In some experiments caspases and PARP inhibitors were used. Caspase-1 inhibitor, VX-765; Caspase-3 inhibitor, Z-DEVD (Selleckchem, Houston, TX, USA); Pan caspase inhibitor, Z-VAD (Tocris Bioscience, Bristol, UK); PARP-1 and -2 inhibitor, ABT-888 and a novel PARP inhibitor, AZD2461 (Selleckchem) were added right before induction of UOS.
3
Protein Purification and Inhibitor Treatments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Clarity Western ECL Substrate was from Bio-Rad (California, USA; #170-5061), and Protein A–Sepharose was from Roche (Milan, Italy; #CL4B 17-0780-01). Digitonin, bafilomycin A1, brefeldin-A, rapamycin and trehalose were purchased from Sigma-Aldrich, Milan, Italy. LY-294002 and Z-VAD were purchased from Tocris Biosciences (UK).
4
Apoptosis Detection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
D-Luciferin was purchased from Perkin Elmer, glosensor (Promega), Z-VAD (Tocris Bioscience), all the other chemicals and reagents required for cell culture including antibiotics, fetal bovine serum (FBS), media were obtained from Invitrogen. Annexin V-PI apoptosis detection kit (BD Biosciences), APC anti-human CD124 (IL-4Rα) and IgG isotype antibody were procured from Biolegend.
5
Elvitegravir-Induced Erythrocyte Alterations
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh Li-Heparin-anticoagulated blood samples were kindly provided by the blood bank of the University of Tübingen. The study is approved by the ethics committee of the University of Tübingen (184/2003 V). The blood was centrifuged at 120 x g for 20 min at 21 °C and the platelets and leukocytes-containing supernatant was disposed. Erythrocytes were incubated for 48 hours at 37°C in vitro at a hematocrit of 0.4% in Ringer solution containing (in mM) 125 NaCl, 5 KCl, 1 MgSO 4 , 32 N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (HEPES; pH 7.4), 5 glucose, and 1 CaCl 2 . Where indicated, erythrocytes were exposed for 48 hours to Elvitegravir (MedChem Express, Princeton, USA). To test for an involvement of p38 kinase, erythrocytes were exposed for 48 hours to a combination of Elvitegravir and p38 kinase inhibitor SB203580 (Tocris bioscience, Bristol, UK). To test for an involvement of caspases, erythrocytes were exposed for 48 hours to a combination of Elvitegravir and pancaspase inhibitor zVAD (Tocris bioscience, Bristol, UK).
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!