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.
Please mouse over
Product Details
| Applications: | Flow Cytometry, Fluorescence microscopy, Fluorescent detection
Microplate
|
|
| Application Notes: | This kit has been specifically designed for monitoring of protein aggregate formation in solution. |
|
| 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
For ENZ-51023-KP002: 2 x 96-well tests or 70 flow cytometry tests |
|
| 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. |
|
| Shipping: | Shipped on Dry Ice |
|
| Short Term Storage: | -20°C |
|
| Long Term Storage: | -80°C |
|
| Contents: | PROTEOSTAT® detection reagent
PROTEOSTAT® positive control
PROTEOSTAT® negative control
10X PROTEOSTAT® assay buffer |
|
| Technical Info/Product Notes: | Application Notes:
Rapid Detection and Characterization of Protein Aggregates by Flow Cytometry
Particle analysis of therapeutic protein formulations with ImageStreamX® Imaging Flow Cytometry and the PROTEOSTAT® Protein Aggregation Assay
Prediction of Aggregation Propensity and Monitoring of Aggregation of Antibody-Drug Conjugates (ADC) using ProteoStat® Reagents
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:
PROTEOSTAT® Cited Samples |
|
| Regulatory Status: | RUO - Research Use Only |
|
Product Literature References
Cell-Instructive Surface Gradients of Photoresponsive Amyloid-like Fibrils: A.M. Bender, et al.; ACS Biomater. Sci. Eng. 7, 4798 (2021), 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;
Endoplasmic reticulum chaperone calmegin is upregulated in aldosterone-producing adenoma and associates with aldosterone production: K. Itcho, et al.; Hypertension 75, 492 (2020), 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;
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
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;
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
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
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;
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
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
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
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;
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;
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;
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
Interfacial dilatational deformation accelerates particle formation in monoclonal antibody solutions: G.L. Lin, et al.; Soft Matter 12, 3293 (2016), Abstract;
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;
A screening methodology for purifying proteins with aggregation problems: M. Lebendiker, et al.; Methods Mol. Biol. 1258, 261 (2015), 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;
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;
WALTZ-DB: a benchmark database of amyloidogenic hexapeptides: J. Beerten, et al.; Bioinformatics 31, 1698 (2015), 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
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;
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
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;
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
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
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;
High sensitivity luminescence nanoparticle assay for the detection of protein aggregation: S. Pihlasalo, et al.; Anal. Chem. 83, 1163 (2011), Abstract;
General Literature References
Emerging analytical technologies for biotherapeutics development: R. Krishnamurthy et al.; Bioprocess International 6 (5), 32 (2008),
Aggregation analysis of therapeutic proteins, part 2: T. Arakawa et al.; Bioprocess International 5 (4), 36 (2007),
Aggregation analysis of therapeutic proteins, part 3: T. Arakawa et al.; Bioprocess International 5(10), 52 (2007),
Aggregation analysis of therapeutic proteins, part 1: T. Arakawa, et al.; Bioprocess International 4 (10), 32 (2006),
Structure of the cross-beta spine of amyloid-like fibrils: R. Nelson et al.; Nature 435, 773 (2005), Abstract;
化工儀器網(wǎng)