Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA

R&D Systems | Catalog # DYC1935-2

R&D Systems
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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
View all HIF-1 alpha/HIF1A ELISA Kits »

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

Please see the product datasheet

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

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.
HIF-1 alpha, as Quantified by the Human/Mouse Total HIF-1 alpha DuoSet IC ELISA

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

The hypoxia-inducible transcription factor 1 alpha (HIF-1 alpha/HIF1A) is the regulated member of the transcription factor heterodimer HIF-1 alpha/beta. HIF-1 alpha/HIF1A binds to hypoxia-response elements (HREs) in the promoters of many genes involved in the adaptation to an environment of insufficient oxygen or hypoxia. Hypoxic tissue environments occur in vascular and pulmonary diseases as well as cancer, which illustrates the broad impact of gene regulation by HIF1A.

Long Name

Hypoxia Inducible Factor 1 Subunit Alpha

Alternate Names

BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8

Entrez Gene IDs

3091 (Human); 15251 (Mouse); 29560 (Rat)

Gene Symbol

HIF1A

Additional HIF-1 alpha/HIF1A Products

Product Documents for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA

For research use only

Citations for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA

Customer Reviews for Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA (6)

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  • Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
    Name: Anonymous
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 05/22/2023
    Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA DYC1935-2
  • Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA
    Name: Anonymous
    Sample Tested: cell supernatant
    Verified Customer | Posted 05/17/2023
    Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA DYC1935-2
  • Human/Mouse Total HIF-1 alpha DuoSet IC ELISA
    Name: Anonymous
    Sample Tested: Tumor cell lyastes
    Verified Customer | Posted 08/01/2019
    Whole tumor lysate was acquired and protein quantified using BCA. 100 ug protein was added to each well from each sample. ELISA was run as per protocol.
    Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA DYC1935-2
  • Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 03/12/2019
  • Human/Mouse Total HIF-1 alpha DuoSet IC ELISA
    Name: Sophie Millar
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 01/16/2019
    Human/Mouse Total HIF-1 alpha/HIF1A DuoSet IC ELISA DYC1935-2
  • Name: Anonymous
    Sample Tested: Plasma
    Verified Customer | Posted 11/10/2018

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