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Aggregate Detection
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
Would membrane proteins work with ProteoStat® Thermal Shift Stability Assay Kit, ENZ-51027, and ProteoStat® Detection Reagent in ProteoStat® Protein Aggregation Assay, ENZ-51023?
We have not tested the ProteoStat® Protein Aggregation Assay, ENZ-51023, or ProteoStat® Thermal Shift Stability Assay Kit, ENZ-51027, with membrane…
Tue, Jul 18, 2023
What are the main advantages of working with ProteoStat® Thermal Shift Stability Assay Kit, ENZ-51027?
The ProteoStat® Thermal Shift Stability Assay kit works over a wide temperature, pH and ionic strength range and is compatible…
Tue, Jul 18, 2023
What does ProteoStat® dye bind to and what is the mechanism of detection of protein aggregation?
The molecular rotor dye is non-fluorescent due to free rotation around a central carbon-carbon single bond separating different aromatic portions of…
Tue, Jul 18, 2023
Are there any limitations regarding particle size of the aggregates that can be detected with the ProteoStat® dye?
The ProteoStat® dye detects in the subvisible to visible range from ~ 50 µm to 1 mm.
Tue, Jul 18, 2023
Two peaks with ProteoStat® Thermal Shift Stability Assay kit, ENZ-51027, were observed. What is the explanation?
One possible explanation for the two peaks is the presence of two proteins in the sample. If the test protein contains…
Tue, Jul 18, 2023
If the test protein is a variant with near identical amino acid sequence with the exception of one residue, is one peak expected ?
There may be a possibility of different conformations of the same protein aggregating at different temperatures. The sample could have…
Tue, Jul 18, 2023