Human Total HSP60 DuoSet IC ELISA

Name: Human Total HSP60 DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC1800-2
Rating: 5 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # DYC1800-2

Citations (3)

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Key Product Details

Assay Type:	Solid Phase Sandwich ELISA
Assay Range:	625 - 40,000 pg/mL
Sample Types:	Cell Lysates
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

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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 for Human Total HSP60 DuoSet IC ELISA

Assay Format	96-well strip plate (sold separately)
Sample Volume Required	Cell lysates (100 µL)
Reactivity	Human
Specificity	Please see the product datasheet
Detection Method	Colorimetric ELISA - 450nm (TMB)
Conjugate	Biotin
Label	HRP
Measurement	Quantitative ELISA
Sufficient Materials	Kits available for two, five, or fifteen 96-well plates*

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

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

Scientific Data Examples 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.

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

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.

Item	Value
Long Name	Heat Shock Protein 60
Entrez Gene IDs	3329 (Human); 110907 (Mouse); 63868 (Rat)
Alternate Names	60 kDa chaperonin, Chaperonin 60, GROEL, HLD4, HSP-60, HSP60, HSP60SPG13, HSP65, HSPD1, Heat shock protein 60, HuCHA60, Mitochondrial matrix protein P1,60 kDa heat shock protein, mitochondrial, P60 lymphocyte protein, SPG13, cpn60, heat shock 60kD protein 1 (chaperonin), heat shock 60kDa protein 1 (chaperonin), heat shock protein 65, short heat shock protein 60 Hsp60s1, spastic paraplegia 13 (autosomal dominant)

Citations for Human Total HSP60 DuoSet IC ELISA (3)

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Citations are publications that use Bio-Techne products. Selected citations for Human Total HSP60 DuoSet IC ELISA include:

Species: Human
Sample Types: Serum

M Wiersma et al. (2020), Cell-Free Circulating Mitochondrial DNA: A Potential Blood-Based Marker for Atrial Fibrillation Cells, 9(5).
PMID: 32397106
Species: Ovine
Sample Types: BALF

(2016), Distending Pressure Did Not Activate Acute Phase or Inflammatory Responses in the Airways and Lungs of Fetal, Preterm Lambs PLoS ONE, 11(7):e0159754.
PMID: 27463520
Species: Ovine
Sample Types: BALF

Hillman N et al. (2014), Sustained inflation at birth did not alter lung injury from mechanical ventilation in surfactant-treated fetal lambs. PLoS ONE, 9(11):e113473.
PMID: 25419969

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Human Total HSP60 DuoSet IC ELISA

Name: Neda Ghousifam

Sample Tested: Human Renal Epithelial Cells

Verified Customer | Posted 09/09/2018

HSP60 was measured in cell lysates samples.

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Product Documents for Human Total HSP60 DuoSet IC ELISA

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Human Total HSP60 DuoSet IC ELISA

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Complete Your Research

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Summary for Human Total HSP60 DuoSet IC ELISA

Product Specifications for Human Total HSP60 DuoSet IC ELISA

Kit Contents for Human Total HSP60 DuoSet IC ELISA

Other Reagents Required

Scientific Data Examples for Human Total HSP60 DuoSet IC ELISA

Preparation and Storage

Background: HSP60

Citations for Human Total HSP60 DuoSet IC ELISA (3)

Customer Reviews for Human Total HSP60 DuoSet IC ELISA (1)

Customer Images

Human Total HSP60 DuoSet IC ELISA

Product Documents for Human Total HSP60 DuoSet IC ELISA

Product Datasheets

COA

Certificate of Analysis

SDS

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