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Mouse HIF-1α(Hypoxia Inducible Factor 1 Alpha) ELISA Kit

Citations(10)Uniprot : Q61221

Cat.No.:E-EL-M0687
Reactivity: Mouse

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	125.00-8000 pg/mL
Sensitivity	75.00 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Mouse HIF-1α in samples. No significant cross-reactivity or interference between Mouse HIF-1α and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Mouse HIF-1α concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse HIF-1α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse HIF-1α and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse HIF-1α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse HIF-1α. You can calculate the concentration of Mouse HIF-1α in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse HIF-1α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse HIF-1α were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	387.70	1086.20	2742.50	371.80	1120.30	2531.80
Standard deviation	24.80	46.70	109.70	19.70	58.30	108.90
CV (%)	6.40	4.30	4.00	5.30	5.20	4.30

Recovery

The recovery of Mouse HIF-1α spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	86-100	93
EDTA plasma (n=8)	93-105	99
Cell culture media (n=8)	89-102	95

Linearity

Samples were spiked with high concentrations of Mouse HIF-1α and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: Q61221
Synonyms	HIF1A, HIF-1A, MOP1, PASD8, bHLHe78, basic helix-loop-helix transcription factor
Research Area	Cancer, Cardiovascular, Epigenetics and Nuclear Signaling, Metabolism

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

ADVANCED MATERIALS (2022) IF: 32.086
Supramolecular Nitric Oxide Depot for Hypoxic Tumor Vessel Normalization and Radiosensitization

DOI: 10.1002/adma.202202625

PMID: 35906003
Sample: B16 cell-xenograft
JOURNAL OF INVESTIGATIVE DERMATOLOGY (2021) IF: 8.551
Successful Treatment of Vitiligo with Cold Atmospheric Plasma‒Activated Hydrogel

DOI: 10.1016/j.jid.2021.04.019

PMID: 34029575
Sample: Serum,Tissue homogenate
BIOMEDICINE & PHARMACOTHERAPY (2019) IF: 6.53
Sinomenine mitigates collagen-induced arthritis mice by inhibiting angiogenesis

DOI: 10.1016/j.biopha.2019.108759

PMID: 30856539
Sample: Serum
Journal of Inflammation Research (2021) IF: 6.922
Histone Deacetylase 6 Regulates the Activation of M1 Macrophages by the Glycolytic Pathway During Acute Liver Failure

DOI: 10.2147/JIR.S302391

Sample: liver,RAW264.7 cell
JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
Amelioration of diabetic retinopathy in db/db mice by treatment with different proportional three active ingredients from Tibetan medicine Berberis dictyophylla F

DOI: 10.1016/j.jep.2021.114190

PMID: 33964362
Sample: Serum
JOURNAL OF PROTEOME RESEARCH (2021) IF: 4.466
Uncovering the Metabolic Mechanism of Salidroside Alleviating Microglial Hypoxia Inflammation Based on Microfluidic Chip-Mass Spectrometry

DOI: 10.1021/acs.jproteome.1c00647

PMID: 34851127
Sample: BV2 cell
Stem Cell Research & Therapy (2018) IF: 4.963
LincRNA-p21 promotes mesenchymal stem cell migration capacity and survival through hypoxic preconditioning

DOI: 10.1186/s13287-018-1031-x

PMID: 30359325
Sample: Cell culture supernatant
CEREBROVASCULAR DISEASES (2016) IF: 3.359
MLC901 Favors Angiogenesis and Associated Recovery after Ischemic Stroke in Mice

DOI: 10.1159/000444810

Sample: Tissue homogenate
INFLAMMATION (2018) IF: 2.884
RNAi Silencing of HIF-1α Ameliorates Lupus Development in MRL/lpr Mice

DOI: 10.1007/s10753-018-0815-6

PMID: 30043119
Sample: Serum
BIOMEDICAL AND ENVIRONMENTAL SCIENCES (2019) IF: 1.917
Effects of Simulated Heat Wave and Ozone on High Fat Diet ApoE Deficient Mice

DOI: 10.3967/bes2018.101

Sample: Plasma,Tissue homogenate

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