- A simple, sensitive, homogenous fluorescent assay
- Validated for use with microplate or flow cytometry platform
- Extensively benchmarked with IgG
- Optimize buffers and excipients for protein formulation
- Performs with a wide pH and ionic strength range
- Use with PROTEOSTAT® Protein Aggregation Standards for accurate quantification of aggregated protein in solution.
PROTEOSTAT
® Protein aggregation assay provides a simple, homogenous assay format for monitoring peptide and protein aggregation in solution. This is useful for defining optimal storage formulations for proteins, for screening of compounds that promote or inhibit protein aggregation and potentially for the sensitive measurement of molecular chaperone activity. The assay can be employed to streamline protein processing and formulation optimization procedures. Relative to conventional protein aggregation detection dyes, such as Thioflavin T, the Enzo PROTEOSTAT
® detection reagent can detect aggregates from a broader range of proteins, yields a much brighter signal, provides at least 2 orders of magnitude linear dynamic range and offers superior performance across a broad range of pH values (4~10) and buffer compositions. Sensitivity for this assay is in the sub-micromolar range and as little as 1-5% protein aggregate is detectable in a concentrated protein solution. The assay is capable of providing quantitative analysis of protein aggregation in a robust and high-throughput fashion (Z’ factor score >0.5). Lyophilized native and aggregated protein are provided as negative and positive controls for monitoring changes in protein aggregation status.
PROTEOSTAT
® Protein Aggregation Standards kit (Prod. No.
ENZ-51039) is the
only commercially available protein aggregation standards assay with stabilized, high-quality reference samples for generating trace protein aggregate levels in concentrated monomeric IgG.
Easy to use -
simply add water!
Effective linear dynamic range for antibody aggregate detection using PROTEOSTAT® Detection Reagent compared with Thioflavin T. Relative fluorescence unit values (RFUs) may differ depending upon the microplate reader employed for the analysis.
Dye is immobilized when bound to the aggregate and begins to fluoresce.
Thioflavin T: early prototype dye in the design of PROTEOSTAT® assay which also rotates around a single bond (red arrow) in the absence of protein aggregates.
Figure 2. Flow Cytometry Application: PROTEOSTAT® Dye Mixed with Bodipy Dye (Pyrromethene 546) Differentiates Oil Droplets From True Protein Aggregates. Bodipy vs PROTEOSTAT® fluorescence of IgG aggregates (A), Silicon oil droplets (B) and a mixture of Silicon oil droplets and IgG aggregates (C).
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Product Details
Product Literature References
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Optimization of protein purification and characterization using Thermofluor screens: S. Boivin, et al.; Protein Expr. Pur.
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Correction of both NBD1 energetics and domain interface is required to restore ΔF508 CFTR folding and function: W.M. Rabeh, et al.; Cell
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Detection of α-synuclein amyloidogenic aggregates in vitro and in cells using light-switching dipyridophenazine ruthenium(II) complexes: N.P. Cook, et al.; J. Am. Chem. Soc.
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p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions: Y. Watanabe, et al.; PLoS One
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Raster image correlation spectroscopy as a novel tool for the quantitative assessment of protein diffusional behaviour in solution: Z. Hamrang, et al.; J. Pharm. Sci.
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