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

Cat.No.:E-EL-R0513
Reactivity: Rat

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48T
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96T*5
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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	7.81-500 pg/mL
Sensitivity	4.69 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat HIF-1α in samples. No significant cross-reactivity or interference between Rat 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 Rat 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 Rat 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 Rat 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 Rat 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 Rat HIF-1α. You can calculate the concentration of Rat 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 Rat 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 Rat 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)	23.60	50.30	194.20	22.50	49.60	180.80
Standard deviation	1.30	2.70	6.20	1.20	2.10	6.30
CV (%)	5.51	5.37	3.19	5.33	4.23	3.48

Recovery

The recovery of Rat 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)	95-108	102
EDTA plasma (n=8)	87-99	93
Cell culture media (n=8)	85-99	90

Linearity

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

Target Information

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

Antioxidants (2022) IF: 7.675
Effect of Empagliflozin on Thioacetamide-Induced Liver Injury in Rats: Role of AMPK/SIRT-1/HIF-1α Pathway in Halting Liver Fibrosis

DOI: 10.3390/antiox11112152

Sample: liver
EUROPEAN JOURNAL OF PHARMACOLOGY (2023) IF: 5.195
Linagliptin counteracts rotenone’s toxicity in non-diabetic rat model of Parkinson’s disease: Insights into the neuroprotective roles of DJ-1, SIRT-1/Nrf-2 and implications of HIF1-α

DOI: 10.1016/j.ejphar.2023.175498

Sample: midbrain,striatum
CELL TRANSPLANTATION (2022) IF: 4.064
Exosomes Derived From Mesenchymal Stem Cells Pretreated With Ischemic Rat Heart Extracts Promote Angiogenesis via the Delivery of DMBT1:

DOI: 10.1177/09636897221102898

Sample: heart tissue
Drug Design Development and Therapy (2023) IF: 4.319
Anti-Arthritic Effect of Edaravone Against Complete Freund Adjuvant Induced Arthritis via Osteoclast Differentiation and HIF-1α–VEGF–ANG-1 Axis

DOI: 10.2147/DDDT.S391606

Sample: serum
JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
Investigation on the Q-markers of Bushen Huoxue Prescriptions for DR treatment based on chemometric methods and spectrum-effect relationship

DOI: 10.1016/j.jep.2021.114800

PMID: 34748867
Sample: eyes
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY (2021) IF: 3.377
Metformin ameliorates bladder dysfunction in a rat model of partial bladder outlet obstruction

DOI: 10.1152/ajprenal.00625.2020

PMID: 33645317
Sample: Tissue homogenate(Bladder)
CELLULAR AND MOLECULAR NEUROBIOLOGY (2020) IF: 3.606
Sulforaphane Induces Glioprotection After LPS Challenge

DOI: 10.1007/s10571-020-00981-5

PMID: 33079284
Sample: Cell lysate
JOURNAL OF FOOD BIOCHEMISTRY (2022) IF: 3.654
Twelve weeks of treadmill exercise training with green tea extract reduces myocardial oxidative stress and alleviates cardiomyocyte apoptosis in aging rat: The emerging role of BNIP3 and HIF-1α/IGFBP3 pathway

DOI: 10.1111/jfbc.14397

PMID: 36069470
Sample: heart tissue
INTERNATIONAL IMMUNOPHARMACOLOGY (2020) IF: 3.943
Alleviation of remote lung injury following liver ischemia/reperfusion: Possible protective role of vildagliptin

DOI: 10.1016/j.intimp.2020.107305

PMID: 33388732
Sample: lung tissue
NAUNYN-SCHMIEDEBERGS ARCHIVES OF PHARMACOLOGY (2019) IF: 2.058
Protective effect of dipeptidyl peptidase-4 inhibitors in testicular torsion/detorsion in rats: a possible role of HIF-1α and nitric oxide

DOI: 10.1007/s00210-019-01765-5

Sample: testicular
Molecular Medicine Reports (2022) IF: 2.952
Reparative effects of chronic intermittent hypobaric hypoxia pre‑treatment on intervertebral disc degeneration in rats

DOI: 10.3892/mmr.2022.12689

PMID: 35315494
Sample: serum
TROPICAL JOURNAL OF PHARMACEUTICAL RESEARCH (2020) IF: 0.504
Effect of penehyclidine hydrochloride on IL-6, TNF-α, HIF- 1α and oxidative stress in lung tissue of young rats with acute lung injury

DOI: 10.4314/tjpr.v19i7.14

Sample: Serum

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