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CELLESTIAL® Assays
Can cell lysates be used to determine protein aggregation with ProteoStat® Aggresome detection reagent?
Working with cell lysates would be challenging, because detergents can cause high background. It is possible to titrate the amount…
Tue, Jul 18, 2023
Can the Proteostat kit be used to study protein aggregation with prokaryote samples?
The dye should work with prokaryotes if membrane fractions are separated. If the membrane fraction is not removed, the dye…
Tue, Jul 18, 2023
Can the Proteostat kit be used to study protein aggregation with gram negative bacteria?
LPS in outer membrane of gram negative bacteria can be problematic. The permeabilizing buffer described in the manual, would remove…
Tue, Jul 18, 2023
Can the FluoForte® Calcium assay kit be used to monitor Ca2+ flux in nicotinic acetylcholine receptors on Flexstation. Can the FluoForte dye monitor Ca2+ in our model system given that FluoForte dye has been validated only in GPCR signaling path
While no specific references are available for application of FluoForte to assay nicotinic receptor, there are numerous publications regarding the…
Tue, Jul 18, 2023
Can calcium mobilization be observed over the course of 1 hour with FluoForte Calcium assay kit? How long can the cells retain the fluorescent dye?
Ligand binding to GPCR generates very fast kinetics for calcium release, with the mobilization complete in less than a minute…
Tue, Jul 18, 2023
Can the cells be trypsinized to determine protein aggregation in cell based assay for testing drug responsiveness?
Trypsin can be used with ProteoStat® Aggresome detection kit. Following lysis, detergent can be added to permeabilize the cells. The…
Tue, Jul 18, 2023
What is a good positive control to study protein aggregation in cells?
MG132 treated cells would serve as positive control to determine background noise compared to signal along with untreated negative control.
Tue, Jul 18, 2023
Would diluting the sample to a lower concentration (e.g. 0.5 mg/mL) change the percentage of aggregate protein?
The percent aggregate (the ratio of aggregate to total protein) will stay the same. However, the concentration of aggregated protein…
Tue, Jul 18, 2023
Is there a step in the protocol of ENZ-51035 at which it is possible to stop and store the samples at 40 C overnight and the protocols can be completed the next day?
It is advisable to stop the protocol if desired before loading the dye. The intensity of the dye decreases over…
Tue, Jul 18, 2023
Can the detection reagent from ProteoStat® Protein Aggregation Assay, ENZ-51023, be substituted with ProteoStat® TS Detection Reagent from the thermal shift stability assay kit, ENZ-51027?
Theoretically, the two dye reagents can be interchanged but this would require some optimization.
Tue, Jul 18, 2023