Human Total HSP60 DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC1800-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
625-40000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human
Human Total HSP60 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
Product Summary for Human Total HSP60 DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total HSP60 in cell lysates. An immobilized capture antibody specific for HSP60 binds both phosphorylated and unphosphorylated HSP60. 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 Total HSP60 DuoSet IC ELISA
Standard Curve
Specificity of Human Total HSP60 DuoSet IC ELISA is Shown by Western Blot.
Lysates prepared from Jurkat human acute T cell leukemia cells were incubated in wells coated with Human Total HSP60 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 an Immobilon-P membrane (Millipore) and immunoblotted with Human Total HSP60 Detection Antibody. Only a single band corresponding to HSP60 was detected. To further determine specificity, recombinant human (rh) HSP27 (Catalog # 1580-HS) and rhHSP70 (Catalog # AP-100) were assayed at 400 ng/mL and did not cross-react or interfere in the assay.
Quantifications of HSP60 in Human Cell Lysates.
Lysates prepared from Md from MCF-7 human breast cancer cells, HeLa human cervical epithelial carcinoma cells and Jurkat human acute T cell leukemia cells were quantified with this DuoSet IC ELISA. The same lysates were also immunoblotted (inset) with Mouse Anti-human HSP60 (Catalog # MAB1800) Antibody. The DuoSet IC ELISA results correlate well with the relative amounts of HSP60 detected by Western blot.Kit Contents for Human Total HSP60 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: HSP60
Long Name
Heat Shock Protein 60
Alternate Names
cpn60, GROEL, HSPD1, SPG13
Gene Symbol
HSPD1
Additional HSP60 Products
Product Documents for Human Total HSP60 DuoSet IC ELISA
Product Specific Notices for Human Total HSP60 DuoSet IC ELISA
For research use only
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