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PROTEOSTAT® PDI assay kit

 
ENZ-51024-KP050 50 tests 217.00 USD
 
ENZ-51024-KP002 2x96 tests 579.00 USD
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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.

Product Specification

Alternative Name:Protein disulfide isomerase
 
Applications:Fluorescent detection
 
Application Notes:This kit provides a simple, homogenous assay for screening modulators of protein disulfide isomerase (PDI) enzymatic activity.
 
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.
 
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.
 
Handling:Protect from light. Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Contents:PROTEOSTAT® PDI detection reagent
PDI (human) (recombinant)
Insulin (Lyophilized)
Bacitracin (Inhibitor Control, Lyophilized)
PBE buffer
Stop reagent
DTT (10mM)
Deionized water
 
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.
 
UniProt ID:P07237 (human)
 
Regulatory Status:RUO - Research Use Only
 

Product Literature References

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;

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