Quantitative Detection of Protein Aggregates from Visible to Subvisible Particles
- 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!
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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).

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.







Product Details
Application |
Flow Cytometry, Fluorescence microscopy, Fluorescent detection, Microplate |
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Application Notes |
This kit has been specifically designed for monitoring of protein aggregate formation in solution. |
Contents |
PROTEOSTAT® detection reagent |
Quality Control |
A sample of PROTEOSTAT® Protein aggregation assay was used to assay (1) 20 µM aggregated lysozyme, (2) 20 µM native lysozyme; and (3) mixtures of 5% aggregate. The Z’ factor is greater than 0.5 and the 5% aggregate signal is greater than 3 standard deviations above the no-aggregate control. |
Quantity |
For ENZ-51023-KP050: 50 tests in a 96 well plate or 16 flow cytometry tests |
Technical Info / Product Notes |
Application Notes: Enzo and PROTEOSTAT are trademarks of Enzo Life Sciences, Inc. Several of Enzo’s products and product applications are covered by US and foreign patents and patents pending. Cited samples: |
Handling & Storage
Use/Stability |
With proper storage, the kit components are stable up to the date noted on the product label. Store kit at -20°C in a non-frost free freezer or -80°C for longer term storage. |
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Short Term Storage |
-20°C |
Long Term Storage |
-80°C |
Shipping |
Dry Ice |
Regulatory Status |
RUO – Research Use Only |
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- Male sex determination maintains proteostasis and extends lifespan of daf-18/PTEN deficient C. elegans: Qu, Z., Zhang, L., et al.; EMBO Rep. 26, 1084 (2025), Abstract
- A superantigen-based MHC class II targeted cancer immunotherapy for the treatment of acute myeloid leukemia: Dolloff, N., Golick, L., et al.; Research Square , (2025)
- Impacts of cytoplasmic p53 aggregates on the prognosis and the transcriptome in lung squamous cell carcinoma: K. Nishitsuji, et al.; Cancer Sci. 115, 2947 (2024), Abstract
- Astragaloside IV reduces mutant Ataxin-3 levels and supports mitochondrial function in Spinocerebellar Ataxia Type 3: Lin, Y., Cheng, W., et al.; Sci. Rep. 14, 25979 (2024), Abstract
- ER-associated degradation adapter Sel1L is required for CD8+ T cell function and memory formation following acute viral infection: Correa-Medero, L. O., Jankowski, S. E., et al.; Cell Rep. 43, 114156 (2024), Abstract
- Exploiting sweet relief for preeclampsia by targeting autophagy-lysosomal machinery and proteinopathy: Huang, Z., Cheng, S., et al.; Exp. Mol. Med. 56, 1206 (2024), Abstract
- Investigating Protein Binding with the Isothermal Ligand-induced Resolubilization Assay: Prout-Holm, R. A., van Walstijn, C. C., et al.; Chembiochem 25, e202300773 (2024), Abstract
- Induction of proteasomal activity in mammalian cells by lifespan-extending tRNA synthetase inhibitors: B.L. Mariner, et al.; Geroscience 46, 1755 (2024), Abstract
- Overexpression of long noncoding RNA DUXAP8 inhibits ER-phagy through activating AKT/mTOR signaling and contributes to preeclampsia: Wei, X. H., Liao, L. Y., et al.; Cell. Mol. Life Sci. 81, 336 (2024), Abstract
- Genome-wide CRISPR screen identifies neddylation as a regulator of neuronal aging and AD neurodegeneration: Saurat, N., Minotti, A. P., et al.; Cell Stem Cell 31, 1162 (2024), Abstract
- The protective effect of erinacine A-enriched Hericium erinaceus mycelium ethanol extract on oxidative Stress–Induced neurotoxicity in cell and Drosophila models of spinocerebellar ataxia type 3: Y.L. Wu, et al.; Free Radic. Biol. Med. 195, 1 (2023), Abstract
- Ubiquitin-like protein MNSFβ regulates glycolysis and promotes cell proliferation with HSC70 assistance: M. Nakamura, et al.; Biochem. Biophys. Rep. 33, 101414 (2023), Abstract
- Protein adduction causes non-mutational inhibition of p53 tumor suppressor: Caspa Gokulan, R., Paulrasu, K., et al.; Cell Rep. 42, 112024 (2023), Abstract
- Comparison of methods generating antibody-epitope conjugates for targeting cancer with virus-specific T cells: W. van der Wulp, et al.; Front. Immunol. 13, 1183914 (2023), Abstract
- Inverse design of viral infectivity-enhancing peptide fibrils from continuous protein-vector embeddings: K. Kaygisiz, et al.; Biomater. Sci. 11, 5251 (2023), Abstract
- Non-functional ubiquitin C-terminal hydrolase L1 drives podocyte injury through impairing proteasomes in autoimmune glomerulonephritis: Reichelt, J., Sachs, W., et al.; Nat. Commun. 14, 2114 (2023), Abstract
- EVA1A regulates hematopoietic stem cell regeneration via ER-mitochondria mediated apoptosis: B. Liu, et al.; Cell Death Dis. 14, 71 (2023), Abstract
- Thiadiazolidinone (TDZD) Analogs Inhibit Aggregation-Mediated Pathology in Diverse Neurodegeneration Models, and Extend C. elegans Life- and Healthspan: S. Kakraba, et al.; Pharmaceuticals 16, 1498 (2023), Abstract
- DNMT2/TRDMT1 gene knockout compromises doxorubicin-induced unfolded protein response and sensitizes cancer cells to ER stress-induced apoptosis: J.A. Grochala, et al.; Apoptosis 28, 166 (2023), Abstract
- Amyloid-like RIP1/RIP3 RHIM Fragments’ Characterization and Application as a Drug Depot: M. Ismail, et al.; Molecules 28, 1480 (2023), Abstract
- Ambra1 haploinsufficiency in CD1 mice results in metabolic alterations and exacerbates age-associated retinal degeneration: Ramírez-Pardo, I., Villarejo-Zori, B., et al.; Autophagy 19, 784 (2023), Abstract
- Quantitative super-resolution imaging of pathological aggregates reveals distinct toxicity profiles in different synucleinopathies: M.J. Morten, et al.; PNAS 119, e2205591119 (2022), Abstract
- The protein aggregation inhibitor YAT2150 has potent antimalarial activity in Plasmodium falciparum in vitro cultures: I.B. Arnaiz, et al.; BMC Biol. 20, 197 (2022), Abstract
- 4-phenylbutyric acid – Identity crisis; can it act as a translation inhibitor?: D. Stein, et al.; Aging Cell 21, e13738 (2022), Abstract
- Ambra1 haploinsufficiency results in metabolic alterations and exacerbates age-associated retinal degeneration: Ramírez-Pardo, I., Villarejo-Zori, B., et al.; bioRxiv , (2022)
- Hypoxia-Reoxygenation Impairs Autophagy-Lysosomal Machinery in Primary Human Trophoblasts Mimicking Placental Pathology of Early-Onset Preeclampsia: Cheng, S., Huang, Z., et al.; Int. J. Mol. Sci. 23, (2022), Abstract
- PERK reprograms hematopoietic progenitor cells to direct tumor-promoting myelopoiesis in the spleen: Liu, M., Wu, C., et al.; J. Exp. Med. 219, (2022), Abstract
- Targeting DNA topoisomerases or checkpoint kinases results in an overload of chaperone systems, triggering aggregation of a metastable subproteome: Huiting, W., Dekker, S. L., et al.; Elife 11, (2022), Abstract
- Surface cysteine to serine substitutions in IL-18 reduce aggregation and enhance activity: J. Saetang, et al.; Peer J. 10, e13626 (2022), Abstract
- Photoinduced Amyloid Fibril Degradation for Controlled Cell Patterning: K. Kaygisiz, et al.; Macromol. Biosci. 0.1002, e2200294 (2022), Abstract
- Protocol for isolation and proteostatic analysis of sub-populations of spermatogenic cells in mouse: Q. Zou, et al.; STAR Protoc. 3, 101398 (2022), Abstract
- Identification of an endoplasmic reticulum proteostasis modulator that enhances insulin production in pancreatic β cells: M. Miyake, et al.; Cell Chem. Biol. 1016, S2451 (2022), Abstract
- Exposure of α-Synuclein Aggregates to Organotypic Slice Cultures Recapitulates Key Molecular Features of Parkinson’s Disease: S. Moudio, et al.; Front. Neurol. 13, 826102 (2022), Abstract
- GrpEL1 regulates mitochondrial unfolded protein response after experimental subarachnoid hemorrhage in vivo and in vitro: C. Ma, et al.; Brain Res. Bull. 181, 97 (2022), Abstract
- Colonization of the Caenorhabditis elegans gut with human enteric bacterial pathogens leads to proteostasis disruption that is rescued by butyrate: Walker, A. C., Bhargava, R., et al.; PLoS Pathog. 17, e1009510 (2021), Abstract
- Novel blood test for early biomarkers of preeclampsia and Alzheimer’s disease: Cheng, S., Banerjee, S., et al.; Sci. Rep. 11, 15934 (2021), Abstract
- Single-nucleus transcriptomic landscape of primate hippocampal aging: Zhang, H., Li, J., et al.; Protein Cell 12, 695 (2021), Abstract
- Cell-Instructive Surface Gradients of Photoresponsive Amyloid-like Fibrils: A.M. Bender, et al.; ACS Biomater. Sci. Eng. 7, 4798 (2021), Abstract
- Proteostasis regulated by testis-specific ribosomal protein RPL39L maintains mouse spermatogenesis: Q. Zou, et al.; iScience 24, 103396 (2021), Abstract
- Rejuvenation of tumour-specific T cells through bispecific antibodies targeting PD-L1 on dendritic cells: L. Liu, et al.; Nat. Biomed. Eng. 5, 1261 (2021), Abstract
- ATP-citrate lyase promotes axonal transport across species: A. Even, et al.; Nat. Commun. 12, 5878 (2021), Abstract
- PERK participates in cardiac valve development via fatty acid oxidation and endocardial-mesenchymal transformation: T. Shimizu, et al.; Sci. Rep. 10, 20094 (2020), Abstract — Full Text
- Endoplasmic reticulum chaperone calmegin is upregulated in aldosterone-producing adenoma and associates with aldosterone production: K. Itcho, et al.; Hypertension 75, 492 (2020), Abstract
- Loss of N-Glycanase 1 Alters Transcriptional and Translational Regulation in K562 Cell Lines: Mueller, W. F., Jakob, P., et al.; G3 (Bethesda) 10, 1585 (2020), Abstract
- The polyphenol quercetin protects from glucotoxicity depending on the aggresome in Caenorhabditis elegans: M. Civelek, et al.; Eur. J. Nutr. 59, 485 (2020), Application(s): Protein aggregation in solution, Abstract
- Monitoring plasma protein aggregation during aging using conformation-specific antibodies and FTIR spectroscopy: S. Magalhaes, et al.; Clin. Chim. Acta 502, 25 (2020), Application(s): Protein aggregation in human plasma samples, Abstract
- Characterization of Protein Aggregates, Silicone Oil Droplets, and Protein-Silicone Interactions using Imaging Flow Cytometry: C. Probst; J. Pharm. Sci. 109, 364 (2020), Abstract
- Evidence for lysosomal biogenesis proteome defect and impaired autophagy in preeclampsia: Nakashima, A., Cheng, S. B., et al.; Autophagy 16, 1771 (2020), Abstract
- Adaptive endoplasmic reticulum stress signalling via IRE1α-XBP1 preserves self-renewal of haematopoietic and pre-leukaemic stem cells: Liu, L., Zhao, M., et al.; Nat. Cell Biol. 21, 328 (2019), Abstract
- Transthyretin amyloid fibril disrupting activities of extracts and fractions from Juglans mandshurica maxim. var. cordiformis (Makino) kitam: N. Chaudhary, et al.; Molecules 24, 500 (2019), Application(s): Aged amyloid fibrils, Abstract — Full Text
- Fibril formation and therapeutic targeting of amyloid-like structures in a yeast model of adenine accumulation: D. Laor, et al.; Nat. Commun. 10, 62 (2019), Abstract — Full Text
- Discovering putative prion-like proteins in Plasmodium falciparum: A computational and experimental analysis: I. Pallares, et al.; Front. Microbiol. 9, 1737 (2018), Application(s): P. falciparum culture, Abstract — Full Text
- High Throughput Differential Scanning Fluorimetry (DSF) Formulation Screening with Complementary Dyes to Assess Protein Unfolding and Aggregation in Presence of Surfactants: S.M. McClure, et al.; Pharm. Res. 35, 81 (2018), Abstract
- The catalytic inactivation of the N-half of human hexokinase 2 and structural and biochemical characterization of its mitochondrial conformation: M.H. Nawaz, et al.; BioSci. Rep. 38, BSR20171666 (2018), Application(s): Aggregation propensity assessed by DSF, Abstract — Full Text
- Caffeic acid and resveratrol ameliorate cellular damage in cell and Drosophila models of spinocerebellar ataxia type 3 through upregulation of Nrf2 pathway: Y.L. Wu, et al.; Free Radic. Biol. Med. 115, 309 (2018), Abstract
- Curcumin Improves Palmitate-Induced Insulin Resistance in Human Umbilical Vein Endothelial Cells by Maintaining Proteostasis in Endoplasmic Reticulum: M. Ye, et al.; Front. Pharmacol. 8, 148 (2017), Abstract — Full Text
- Aggregated transthyretin is specifically packaged into placental nano-vesicles in preeclampsia: M. Tong, et al.; Sci. Rep. 7, 6694 (2017), Application(s): Protein aggregation in extracts from placental extracellular vesicles, Abstract — Full Text
- Trans ε-viniferin is an amyloid-β disaggregating and anti-inflammatory drug in a mouse primary cellular model of Alzheimer’s disease: E. Vion, et al.; Mol. Cell. Neurosci. 88, 1 (2017), Abstract
- Scaffold requirements for periodontal regeneration with enamel matrix derivative proteins: A. Apicella, et al.; Colloids Surf. B Biointerfaces 156, 221 (2017), Application(s): Aggregation of enaml matrix derivative (EMD) proteins, Abstract
- Interfacial dilatational deformation accelerates particle formation in monoclonal antibody solutions: G.L. Lin, et al.; Soft Matter 12, 3293 (2016), Abstract
- Cardiomyocyte-specific human Bcl2-associated anthanogene 3 P209L expression induces mitochondrial fragmentation, Bcl2-associated anthanogene 3: M.T. Quintana, et al.; Am. J. Pathol. 186, 1989 (2016), Application(s): Lysates from cardiac tissues, Abstract — Full Text
- Resveratrol reduces amyloid-beta (Aβ1–42)-induced paralysis through targeting proteostasis in an Alzheimer model of Caenorhabditis elegans: C. Regitz, et al.; Eur. J. Nutr. 55, 741 (2016), Application(s): Protein aggregation in solution, Abstract
- ARD1-mediated Hsp70 acetylation balances stress-induced protein refolding and degradation: J.H. Seo, et al.; Nat. Commun. 7, 12882 (2016), Application(s): , Abstract — Full Text
- BRG1 and BRM SWI/SNF ATPases redundantly maintain cardiomyocyte homeostasis by regulating cardiomyocyte mitophagy and mitochondrial dynamics in vivo: S.J. Bultman, et al.; Cardiovasc. Pathol. 25, 258 (2016), Abstract
- Effects of tau domain-specific antibodies and intravenous immunoglobulin on tau aggregation and aggregate degradation: J.O. Esteves-Villanueva, et al.; Biochemistry 54, 293 (2015), Abstract
- A screening methodology for purifying proteins with aggregation problems: M. Lebendiker, et al.; Methods Mol. Biol. 1258, 261 (2015), Abstract
- WALTZ-DB: a benchmark database of amyloidogenic hexapeptides: J. Beerten, et al.; Bioinformatics 31, 1698 (2015), Abstract
- Metal-mediated protein oxidation: applications of a modified ELISA-based carbonyl detection assay for complex proteins: H. Uehara, et al.; Pharm. Res. 32, 691 (2015), Application(s): Effects of metal-mediated protein oxidation on aggregation, Abstract
- Lysosomal enzyme cathepsin B enhances the aggregate forming activity of exogenous α-synuclein fibrils: A. Tsujimura, et al.; Neurobiol. Dis. 73C, 244 (2014), Application(s): In vitro formation of a-synuclein fibrils and monitoring with ProteoStat® dye, Abstract
- ProteoStat to detect and discriminate intracellular amyloid-like aggregates in Escherichia coli: S.Navarro & S.Ventura; Biotechnol. J 9, 1259 (2014), Abstract
- Amyloid-beta (Aβ1-42)-induced paralysis in Caenorhabditis elegans is reduced by restricted cholesterol supply: C. Regitz, et al.; Neurosci. Lett. 576, 93 (2014), Abstract
- Human Stefin B Role in Cell’s Response to Misfolded Proteins and Autophagy: M. Polajnar, et al.; PLoS One 9, e102500 (2014), Application(s): Proteins aggregation and oxidative stress in stefin B KO astrocytes, Abstract — Full Text
- Optimization of protein purification and characterization using Thermofluor screens: S. Boivin, et al.; Protein Expr. Pur. 91, 192 (2013), Abstract
- Protein quality control acts on folding intermediates to shape the effects of mutations on organismal fitness: S. Bershtein, et al.; Mol. Cell. 49, 133 (2013), Application(s): Propensity to aggregate for dihydrofolate reductase mutants, Abstract
- Increased carbonylation, protein aggregation and apoptosis in the spinal cord of mice with experimental autoimmune encephalomyelitis: A. Dasgupta, et al.; ASN Neuro 5, e00111 (2013), Application(s): Detection of protein aggregation in tissue section using fluorescence microscopy, Abstract — Full Text
- Correction of both NBD1 energetics and domain interface is required to restore ΔF508 CFTR folding and function: W.M. Rabeh, et al.; Cell 148, 150 (2012), Application(s): Aggregation of multi-domain protein: NBD1, Abstract — Full Text
- 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. 101, 2082 (2012), Abstract
- 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. 134, 20776 (2012), Abstract
- p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions: Y. Watanabe, et al.; PLoS One 7, e52868 (2012), Application(s): Protein aggregation of alpha-synuclein, Abstract — Full Text
- Novel Cell- and Tissue-Based Assays for Detecting Misfolded and Aggregated Protein Accumulation Within Aggresomes and Inclusion Bodies: D. Shen, et al.; Cell Biochem. Biophys. 60, 173 (2011), Abstract
- Multiple Aggregates and Aggresomes of C-Terminal Truncated Human αA-Crystallins in Mammalian Cells and Protection by αB-Crystallin: I. Raju, et al.; PLoS One 6, e19876 (2011), Abstract
- High sensitivity luminescence nanoparticle assay for the detection of protein aggregation: S. Pihlasalo, et al.; Anal. Chem. 83, 1163 (2011), Abstract
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