Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
Kit Content
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
- Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Scientific Data
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The HSP-70 Duoset IC ELISA specifically recognizes HSP-70. Specificity was demonstrated by Western blot analysis of the protein bound by the capture antibody supplied in the kit. Lysates prepared from heat-shocked, human Jurkat cells were incubated in wells coated with HSP-70 Duoset IC Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and captured proteins were electrophoresed, transferred to Immubilon membranes and immunoblotted with HSP-70 Duoset IC Detection Antibody. Only a single band corresponding to HSP-70 was detected.
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Absolute amounts of human HSP-70, as quantified by the HSP-70 DuoSet IC ELISA, are consistent with the amounts of HSP-70 determined by qualitative Western blot analysis. Jurkat cells were heat-shocked at 44ºC for 30 mins and allowed to recover for 4 hrs at 37 ºC. Following cell lysis, HSP-70 was quantified with the DuoSet IC. The same lysates were also immunoblotted (inset) with Rb anti-HSP-70 (R&D Systems, Catalog # AF1663) antibody. The DuoSet IC results correlate well with the amounts of HSP-70 detected by Western blot.
Product Datasheets
Preparation and Storage
Background: HSP70/HSPA1A
The heat shock proteins are a highly conserved family of stress response proteins. HSPs function primarily as molecular chaperones, facilitating the folding of other cellular proteins, preventing protein aggregation, or targeting improperly folded proteins to specific degradative pathways. Some HSPs are expressed at low levels under normal physiological conditions but show dramatically increased expression in response to cellular stress, others are constitutively expressed. Specific HSPs play a role in regulating apoptosis by interacting directly with key components of the apoptotic pathway.
Citations for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Biomarkers in Adult-Type Diffuse Gliomas: Elevated Levels of Circulating Vesicular Heat Shock Protein 70 Serve as a Biomarker in Grade 4 Glioblastoma and Increase NK Cell Frequencies in Grade 3 Glioma
Authors: Lennartz, P;Thölke, D;Bashiri Dezfouli, A;Pilz, M;Lobinger, D;Messner, V;Zanth, H;Ainslie, K;Kafshgari, MH;Rammes, G;Ballmann, M;Schlegel, M;Foulds, GA;Pockley, AG;Schmidt-Graf, F;Multhoff, G;
Biomedicines
Species: Human
Sample Types: Serum, Plasma
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The Stability and Anti-Angiogenic Properties of Titanium Dioxide Nanoparticles (TiO2NPs) Using Caco-2 Cells
Authors: OM David, KL Lategan, MF de Cortale, EJ Pool
Biomolecules, 2022-09-21;12(10):.
Species: Human
Sample Types: Cell Lysates
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Intralesional injection of rose bengal augments the efficacy of gemcitabine chemotherapy against pancreatic tumors
Authors: P Innamarato, J Morse, A Mackay, S Asby, M Beatty, J Blauvelt, S Kidd, JE Mullinax, AA Sarnaik, S Pilon-Thom
BMC Cancer, 2021-06-30;21(1):756.
Species: Mouse
Sample Types: Serum
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Effects of Bovine Colostrum with or without Egg on In Vitro Bacterial-Induced Intestinal Damage with Relevance for SIBO and Infectious Diarrhea
Authors: RJ Playford, N Choudhry, P Kelly, T Marchbank
Nutrients, 2021-03-22;13(3):.
Species: Human
Sample Types: Cell Lysates
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Mitoxantrone-Loaded Nanoparticles for Magnetically Controlled Tumor Therapy-Induction of Tumor Cell Death, Release of Danger Signals and Activation of Immune Cells
Authors: T Ratschker, L Egenberger, M Alev, L Zschiesche, J Band, E Schreiber, B Frey, A Derer, C Alexiou, C Janko
Pharmaceutics, 2020-09-27;12(10):.
Species: Human
Sample Types: Cell Culture Supernates
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A tumor-intrinsic PD-L1-NLRP3 inflammasome signaling pathway drives resistance to anti-PD-1 immunotherapy
Authors: B Theivanthi, KS Evans, NC DeVito, MP Plebanek, M Sturdivant, LP Wachsmuth, AK Salama, Y Kang, D Hsu, JM Balko, DB Johnson, M Starr, AB Nixon, A Holtzhause, BA Hanks
J. Clin. Invest., 2020-05-01;0(0):.
Species: Mouse
Sample Types: Plasma
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Differences of the Immune Phenotype of Breast Cancer Cells after Ex Vivo Hyperthermia by Warm-Water or Microwave Radiation in a Closed-Loop System Alone or in Combination with Radiotherapy
Authors: M Hader, DP Savcigil, A Rosin, P Ponfick, S Gekle, M Wadepohl, S Bekeschus, R Fietkau, B Frey, E Schlücker, US Gaipl
Cancers (Basel), 2020-04-27;12(5):.
Species: Human
Sample Types: Cell Culture Supernates
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Radiosensitization of HSF-1 Knockdown Lung Cancer Cells by Low Concentrations of Hsp90 Inhibitor NVP-AUY922
Authors: A Kühnel, D Schilling, SE Combs, B Haller, M Schwab, G Multhoff
Cells, 2019-09-28;8(10):.
Species: Human
Sample Types: Cell Lysates
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Systemic release of heat-shock protein 27 and 70 following severe trauma
Authors: T Haider, E Simader, O Glück, HJ Ankersmit, T Heinz, S Hajdu, LL Negrin
Sci Rep, 2019-07-03;9(1):9595.
Species: Human
Sample Types: Serum
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Therapeutic ultrasound attenuates DSS-induced colitis through the cholinergic anti-inflammatory pathway
Authors: NS Nunes, P Chandran, M Sundby, F Visioli, F da Costa G, SR Burks, AH Paz, JA Frank
EBioMedicine, 2019-06-26;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Chemo-photothermal therapy combination elicits anti-tumor immunity against advanced metastatic cancer
Authors: J Nam, S Son, LJ Ochyl, R Kuai, A Schwendema, JJ Moon
Nat Commun, 2018-03-14;9(1):1074.
Species: Mouse
Sample Types: Tissue Homogenates
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Disrupting the blood-brain barrier by focused ultrasound induces sterile inflammation
Proc. Natl. Acad. Sci. U.S.A, 2016-12-19;0(0):.
Species: Rat
Sample Types: Tissue Homogenates
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Oncolytic Group B Adenovirus Enadenotucirev Mediates Non-apoptotic Cell Death with Membrane Disruption and Release of Inflammatory Mediators
Authors: A Dyer, Y Di, H Calderon, S Illingwort, G Kueberuwa, A Tedcastle, P Jakeman, SL Chia, A Brown, MA Silva, D Barlow, J Beadle, T Hermiston, DJ Ferguson, B Champion, KD Fisher, LW Seymour
Mol Ther Oncolytics, 2016-12-10;4(0):18-30.
Species: Human
Sample Types: Cell Culture Supernates
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Study of Immunomodulatory Effects of Extracellular HSP70 in a Mouse Model of Allergic Airway Inflammation
Authors: A M Sapozhniko
Biochemistry Mosc., 2016-11-01;81(11):1384-1395.
Species: Mouse
Sample Types: BALF
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A novel HSP90 inhibitor with reduced hepatotoxicity synergizes with radiotherapy to induce apoptosis, abrogate clonogenic survival, and improve tumor control in models of colorectal cancer
Authors: L Kinzel, A Ernst, M Orth, V Albrecht, R Hennel, N Brix, B Frey, US Gaipl, G Zuchtriege, CA Reichel, A Blutke, D Schilling, G Multhoff, M Li, M Niyazi, AA Friedl, N Winssinger, C Belka, K Lauber
Oncotarget, 2016-07-12;7(28):43199-43219.
Species: Human
Sample Types: Cell Culture Supernates
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Defining immunological impact and therapeutic benefit of mild heating in a murine model of arthritis.
Authors: Lee C, Kokolus K, Leigh N, Capitano M, Hylander B, Repasky E
PLoS ONE, 2015-03-20;10(3):e0120327.
Species: Mouse
Sample Types: Cell Culture Supernates
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Bacterial lipopolysaccharide augments febrile-range hyperthermia-induced heat shock protein 70 expression and extracellular release in human THP1 cells.
Authors: Tulapurkar M, Ramarathnam A, Hasday J, Singh I
PLoS ONE, 2015-02-06;10(2):e0118010.
Species: Human
Sample Types: Cell Culture Supernates
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Sustained inflation at birth did not alter lung injury from mechanical ventilation in surfactant-treated fetal lambs.
Authors: Hillman N, Kemp M, Miura Y, Kallapur S, Jobe A
PLoS ONE, 2014-11-24;9(11):e113473.
Species: Ovine
Sample Types: BALF
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Mast cell chymase degrades the alarmins heat shock protein 70, biglycan, HMGB1, and interleukin-33 (IL-33) and limits danger-induced inflammation.
Authors: Roy A, Ganesh G, Sippola H, Bolin S, Sawesi O, Dagalv A, Schlenner S, Feyerabend T, Rodewald H, Kjellen L, Hellman L, Abrink M
J Biol Chem, 2013-11-20;289(1):237-50.
Species: Mouse
Sample Types: Cell Culture Supernates
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Toll-like receptor agonists and febrile range hyperthermia synergize to induce heat shock protein 70 expression and extracellular release.
Authors: Gupta A, Cooper Z, Tulapurkar M, Potla R, Maity T, Hasday J, Singh I
J Biol Chem, 2012-12-04;288(4):2756-66.
Species: Human, Mouse
Sample Types: Cell Culture Supernates
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Aberrant stress-induced Hsp70 expression in immune cells in multiple sclerosis.
Authors: Cwiklinska H, Mycko MP, Szymanska B
J. Neurosci. Res., 2010-11-01;88(14):3102-10.
Species: Human
Sample Types: Cell Lysates
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Regulation of HSP70 on activating macrophages using PDT-induced apoptotic cells.
Authors: Zhou F, Xing D, Chen WR
Int. J. Cancer, 2009-09-15;125(6):1380-9.
Species: Mouse
Sample Types: Cell Culture Supernates
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Serum level of soluble 70-kD heat shock protein is associated with high mortality in patients with colorectal cancer without distant metastasis.
Authors: Kocsis J, Madaras B, Toth EK, Fust G, Prohaszka Z
Cell Stress Chaperones, 2009-07-04;15(2):143-51.
Species: Human
Sample Types: Serum
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Heat shock protein 70 (HSP70) induces cytotoxicity of T-helper cells.
Authors: Figueiredo C, Wittmann M, Wang D, Dressel R, Seltsam A, Blasczyk R, Eiz-Vesper B
Blood, 2008-11-18;113(13):3008-16.
Species: Human
Sample Types: Cell Culture Supernates
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MAPK pathway mediates EGR-1-HSP70-dependent cigarette smoke-induced chemokine production.
Authors: Li CJ, Ning W, Matthay MA, Feghali-Bostwick CA, Choi AM
Am. J. Physiol. Lung Cell Mol. Physiol., 2007-05-01;292(5):L1297-303.
Species: Human
Sample Types: Cell Culture Supernates
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