Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA

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Standard Curve
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Product Details
Citations (24)
Supplemental Products
Reviews (3)

Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Length
4 hours 40 mins (after plate preparation)
Sample Volume Required
Cell lysates (100 µL)
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total HSP70 / HSPA1A in cell lysates. An immobilized capture antibody specific for HSP70 / HSPA1A binds both phosphorylated and unphosphorylated HSP70 / HSPA1A. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

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

Standard Curve

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.

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

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Long Name:
Heat Shock Protein 70
Entrez Gene IDs:
3303 (Human); 15511 (Mouse); 24472 (Rat)
Alternate Names:
dnaK-type molecular chaperone HSP70-1; FLJ54303; FLJ54370; FLJ54392; FLJ54408; FLJ75127; Heat shock 70 kDa protein 1/2; heat shock 70 kDa protein 1A/1B; heat shock 70kD protein 1A; heat shock 70kDa protein 1A; heat shock-induced protein; HSP70; HSP70.1/HSP70.2; HSP70-1; HSP70-1/HSP70-2; HSP70-1A; HSP70I; HSP72; HSPA1; HSPA1A; HSPA1B

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.

24 Citations: Showing 1 - 10
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  1. 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;12(10):.
    Species: Human
    Sample Types: Cell Lysates
  2. 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;13(3):.
    Species: Human
    Sample Types: Cell Lysates
  3. 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;21(1):756.
    Species: Mouse
    Sample Types: Serum
  4. 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;12(10):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. 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;0(0):.
    Species: Mouse
    Sample Types: Plasma
  6. 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;12(5):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. 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;9(1):9595.
    Species: Human
    Sample Types: Serum
  8. 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;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. 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;8(10):.
    Species: Human
    Sample Types: Cell Lysates
  10. 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;9(1):1074.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. 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, 2017;4(0):18-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. 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;7(28):43199-43219.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Study of Immunomodulatory Effects of Extracellular HSP70 in a Mouse Model of Allergic Airway Inflammation
    Authors: A M Sapozhniko
    Biochemistry Mosc., 2016;81(11):1384-1395.
    Species: Mouse
    Sample Types: BALF
  14. Disrupting the blood-brain barrier by focused ultrasound induces sterile inflammation
    Proc. Natl. Acad. Sci. U.S.A, 2016;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  15. 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;10(2):e0118010.
  16. 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;10(3):e0120327.
  17. 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, 2014;289(1):237-50.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. 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;9(11):e113473.
    Species: Ovine
    Sample Types: BALF
  19. 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, 2013;288(4):2756-66.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 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, 2010;15(2):143-51.
    Species: Human
    Sample Types: Serum
  21. Aberrant stress-induced Hsp70 expression in immune cells in multiple sclerosis.
    Authors: Cwiklinska H, Mycko MP, Szymanska B
    J. Neurosci. Res., 2010;88(14):3102-10.
    Species: Human
    Sample Types: Cell Lysates
  22. 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, 2009;113(13):3008-16.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Regulation of HSP70 on activating macrophages using PDT-induced apoptotic cells.
    Authors: Zhou F, Xing D, Chen WR
    Int. J. Cancer, 2009;125(6):1380-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  24. 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;292(5):L1297-303.
    Species: Human
    Sample Types: Cell Culture Supernates


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Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
By Anonymous on 09/10/2018
Sample Tested: Human Renal Epithelial Cells

Human Renal Epithelial Cell Lysates

Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
By Revathi Shanmugasundaram on 02/06/2018
Sample Tested: Serum-free Cell Culture Supernates

Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
By Anonymous on 05/02/2017
Sample Tested: Peritoneal macrophages