- Homogenous, mix-and-read assay for 96-well and 384-well plates
- Widely cited
The PROTEOSTAT® PDI assay kit provides a simple, homogenous assay for screening modulators of PDI enzymatic activity. This is accomplished by monitoring PDI-catalyzed reduction of insulin in the presence of Dithiothreitol (DTT), resulting in the formation of insulin aggregates which then bind avidly to the red-emitting fluorogenic PROTEOSTAT® PDI detection dye. Relative to analogous turbidimetric assays of PDI activity, the fluorescence-based assay provides a vastly improved assay signal window, improved lower detection limit, and superior Z’-score (>0.8). Intra-plate and inter-plate CVs using the assay are typically 3-5%. Human recombinant PDI enzyme (EC 5.3.4.1) and the PDI inhibitor, bacitracin, are included in the kit, as well as all reagents necessary for monitoring changes in PDI activity. The PROTEOSTAT® PDI assay kit is capable of providing a quantitative readout of PDI enzymatic activity in a robust and high-throughput fashion and can be applied to identification of PDI inhibitors from chemical libraries. It is useful in the research of oxidative damage, ER stress and unfolded protein response (UPR).
Product Details
| Alternative Name: | Protein disulfide isomerase |
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| Applications: | Fluorescent detection
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| Application Notes: | This kit provides a simple, homogenous assay for screening modulators of protein disulfide isomerase (PDI) enzymatic activity. |
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| Quality Control: | Using the procedure described in the manual, a sample PROTEOSTAT® PDI assay kit is used to assay the following: (1) Insulin with PDI; (2) Insulin without PDI; and (3) Insulin with PDI and bacitracin. The Z’-factor is greater than 0.8. PDI is inhibited by at least 50% with bacitracin. |
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| 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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| Handling: | Protect from light. Avoid freeze/thaw cycles. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Contents: | PROTEOSTAT® PDI detection reagent
PDI (human) (recombinant)
Insulin (Lyophilized)
Bacitracin (Inhibitor Control, Lyophilized)
PBE buffer
Stop reagent
DTT (10mM)
Deionized water |
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| Technical Info/Product 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. |
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| UniProt ID: | P07237 (human) |
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
Melanoma RBPome identification reveals PDIA6 as an unconventional RNA-binding protein involved in metastasis: N.M. Farràs, et al.; Nucleic Acids Res.
50, 8207 (2022),
Abstract;
Thioredoxin interacting protein protects mice from fasting induced liver steatosis by activating ER stress and its downstream signaling pathways: H. Miyahara, et al.; Sci. Rep.
12, 4819 (2022),
Abstract;
Thioredoxin reductase is a major regulator of metabolism in leukemia cells: S. Karunanithi, et al.; Oncogene
40, 5236 (2021),
Abstract;
Molecular docking‐assisted screening reveals tannic acid as a natural protein disulphide isomerase inhibitor with antiplatelet and antithrombotic activities: L. Ren, et al.; J. Cell. Mol. Med.
24, 14257 (2020),
Abstract;
Full Text
PDI knockdown inhibits seizure activity in acute seizure and chronic epilepsy rat models via S-nitrosylation-independent thiolation on NMDA receptor: A.R. Jeon, et al.; Front. Cell. Neurosci.
12, 438 (2018),
Application(s): Hippocampal tissue lysates,
Abstract;
Full Text
The thiosemicarbazone Me2NNMe2 induces paraptosis by disrupting the ER thiol redox homeostasis based on protein disulfide isomerase inhibition: S. Hager, et al.; Cell Death Dis.
9, 1052 (2018),
Abstract;
Full Text
Tissue Factor Coagulant Activity is Regulated by the Plasma Membrane Microenvironment: Y. Yu, et al.; Thromb. Haemost.
118, 990 (2018),
Application(s): In-vitro activity assay,
Abstract;
Curcumin and Curcuma longa L. extract ameliorate lipid accumulation through the regulation of the endoplasmic reticulum redox and ER stress: H.Y. Lee, et al.; Sci. Rep.
7, 6513 (2017),
Application(s): PDI assay of tissue lysates,
Abstract;
Full Text
Nox4 regulates the eNOS uncoupling process in aging endothelial cells: H.Y. Lee, et al.; Free Radic. Biol. Med.
113, 26 (2017),
Application(s): PDI assay of HUVEC cell lysates,
Abstract;
PDI regulates seizure activity via NMDA receptor redox in rats: J.Y. Kim, et al.; Sci. Rep.
7, 42491 (2017),
Application(s): Hippocampal tissue lysates,
Abstract;
Full Text
Protein disulfide isomerase regulates renal AT1 receptor function and blood pressure in rats: X. Wang, et al.; Am. J. Physiol. Renal. Physiol.
313, F461 (2017),
Application(s): Renal tissue lysates,
Abstract;
Full Text
Requirement of zinc transporter SLC39A7/ZIP7 for dermal development to fine-tune endoplasmic reticulum function by regulating protein disulfide isomerase: B.H. Bin, et al.; J. Invest. Dermatol.
137, 1682 (2017),
Application(s): ER fractions from murine osteoblasts,
Abstract;
Succination of protein disulfide isomerase links mitochondrial stress and endoplasmic reticulum stress in the adipocyte during diabetes: A.M. Manuel, et al.; Antioxid. Redox Signal.
27, 1281 (2017),
Application(s): 3T3-L1 adipocytes and adipose tissue lysates,
Abstract;
Full Text
Adverse outcomes associated with cigarette smoke radicals related to damage to protein disulfide isomerase: H. Kenche, et al.; J. Biol. Chem.
291, 4763 (2016),
Abstract;
Adverse Outcomes Associated with Cigarette Smoke Radicals Related to Damage to Protein-disulfide Isomerase: H. Kenche, et al.; J. Biol. Chem.
291, 4763 (2016),
Application(s): in-vitro assay with recombinant PDI,
Abstract;
Full Text
Role of two sequence motifs of mesencephalic astrocyte-derived neurotrophic factor in its survival-promoting activity: K. Mätlik, et al.; Cell Death Dis.
6, e2032 (2015),
Abstract;
Full Text
Methods of measuring protein disulfide isomerase activity: a critical overview: M.M. Watanabe, et al.; Front. Chem.
2, 73 (2014),
Application(s): Review on available assays to measure PDI activity,
Abstract;
Full Text
N-octanoyl dopamine treatment of endothelial cells induces the unfolded protein response and results in hypometabolism and tolerance to hypothermia: E. Stamellou, et al.; PLoS One
9, e99298 (2014),
Abstract;
Full Text
The protein disulfide isomerases PDIA4 and PDIA6 mediate resistance to cisplatin-induced cell death in lung adenocarcinoma: G. Tufo, et al.; Cell Death Differ.
21, 685 (2014),
Application(s): Evaluation of PDI activity in whole extracts and ER fractions,
Abstract;
Rapid activation of monocyte tissue factor by antithymocyte globulin is dependent on complement and protein disulfide isomerase: F. Langer, et al.; Blood
121, 2324 (2013),
Abstract;
Oxidative processing of latent Fas in the endoplasmic reticulum controls the strength of apoptosis: V. Anathy, et al.; Mol. Cell. Biol.
32, 3464 (2012),
Abstract;
General Literature References
Attachment and entry of Chlamydia have distinct requirements for host protein disulfide isomerase: S. Abromaitis & R.S. Stephens; PLoS Pathog.
5, e1000357 (2009),
Abstract;
Protein disulfide isomerase (PDI) associates with NADPH oxidase and is required for phagocytosis of Leishmania chagasi promastigotes by macrophages: C. X. Santos, et al.; J. Leukoc. Biol.
86, 989 (2009),
Abstract;
The proteostasis boundary in misfolding diseases of membrane traffic: D.M. Hutt, et al.; FEBS Lett.
583, 2639 (2009),
Abstract;
Increasing melanoma cell death using inhibitors of protein disulfide isomerases to abrogate survival responses to endoplasmic reticulum stress: P.E. Lovat, et al.; Cancer Res.
68, 5363 (2008),
Abstract;
Targeting homeostatic mechanisms of endoplasmic reticulum stress to increase susceptibility of cancer cells to fenretinide-induced apoptosis: the role of stress proteins ERdj5 and ERp57: M. Corazzari, et al.; Br. J. Cancer
96, 1062 (2007),
Abstract;
Coupling endoplasmic reticulum stress to the cell death program: R.V. Rao, et al.; Cell Death Differ.
11, 372 (2004),
Abstract;
Protein-disulfide isomerase-mediated reduction of two disulfide bonds of HIV envelope glycoprotein 120 occurs post-CXCR4 binding and is required for fusion: R. Barbouche, et al.; J. Biol. Chem.
278, 3131 (2003),
Abstract;
Protein disulfide isomerase, a component of the estrogen receptor complex, is associated with Chlamydia trachomatis serovar E attached to human endometrial epithelial cells: C.H. Davis, et al.; Infect. Immun.
70, 3413 (2002),
Abstract;
Protein-disulfide isomerase-mediated reduction of the A subunit of cholera toxin in a human intestinal cell line: P.A. Orlandi; J. Biol. Chem.
272, 4591 (1997),
Abstract;
Acceleration of reactivation of reduced bovine pancreatic ribonuclease by a microsomal system from rat liver: R.F. Goldberger, et al.; J. Biol. Chem.
238, 628 (1963),
Abstract;
