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.
Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
R&D Systems | Catalog # DYC1663-2
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
156-10000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human, Mouse, Rat
Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA 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
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Product Summary for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
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 Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
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.
Kit Contents for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
- 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
Preparation and Storage
Shipping
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
Long Name
Heat Shock Protein 70
Alternate Names
HSP70-1A, HSP72, HSPA1A
Gene Symbol
HSPA1A
Additional HSP70/HSPA1A Products
Product Documents for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
Product Specific Notices for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
For research use only
Related Research Areas
Citations for Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA
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Sample Tested: Human Renal Epithelial CellsVerified Customer | Posted 09/10/2018Human Renal Epithelial Cell Lysates
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Sample Tested: Serum-free Cell Culture SupernatesVerified Customer | Posted 02/06/2018
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Sample Tested: Peritoneal macrophagesVerified Customer | Posted 05/02/2017
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
