Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
R&D Systems, part of Bio-Techne | Catalog # DYC1935-2
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
125-8000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human, Mouse
Human/Mouse Total HIF-1 alpha/HIF1A 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/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total HIF-1 alpha in cell lysates. An immobilized capture antibody specific for HIF-1 alpha binds both phosphorylated and unphosphorylated HIF-1 alpha. 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 Total HIF-1 alpha/HIF1A DuoSet IC ELISA
Standard Curve
Specificity of Human/Mouse HIF-1 alpha DuoSet IC ELISA is shown by Western Blot.
Lysates prepared from NIH-3T3 mouse embryonic fibroblast cells, untreated or treated with 150 µM of CoCl2, were incubated in wells coated with Human/Mouse Total HIF-1 alpha Capture Antibody. Unbound material was removed by washing, and bound material was solubilized in SDS gel sample buffer. Captured proteins were electrophoresed, transferred to PVDF membranes and immunoblotted with the Human/Mouse Total HIF-1 alpha Detection Antibody.
Amounts of HIF-1 alpha, as Quantified by the Human/Mouse Total HIF-1 alpha DuoSet IC ELISA, are Consistent with the Relative Amounts of HIF-1 alpha Determined by Qualitative Western Blot Analysis.
Lysates were prepared from MCF-7 human breast cancer cells, untreated or treated with 150 µM CoCl2. HIF-1 alpha was measured with the Human/Mouse Total HIF-1 alpha DuoSet IC ELISA and the same lysates were immunoblotted (inset) with Goat Anti-HIF-1 alpha Antibody (Catalog # AF1935). The DuoSet IC ELISA results correlate with the relative amounts of HIF-1 alpha detected by Western blot.Kit Contents for Human/Mouse Total HIF-1 alpha/HIF1A 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: HIF-1 alpha/HIF1A
Long Name
Hypoxia Inducible Factor 1 Subunit Alpha
Alternate Names
BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8
Gene Symbol
HIF1A
Additional HIF-1 alpha/HIF1A Products
Product Documents for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
Product Specific Notices for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
For research use only
Loading...
Loading...
Loading...