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Rat αFP(Alpha-Fetoprotein) ELISA Kit

Cat.No.:E-EL-R0153
Reactivity: Rat

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat αFP in samples. No significant cross-reactivity or interference between Rat αFP 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 αFP 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 αFP. 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 αFP 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 αFP, 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 αFP. You can calculate the concentration of Rat αFP 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 αFP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat αFP 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 (ng/mL)	1.10	3.00	9.30	1.00	3.30	8.50
Standard deviation	0.10	0.20	0.40	0.10	0.10	0.40
CV (%)	9.09	6.67	4.30	10.00	3.03	4.71

Recovery

The recovery of Rat αFP 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)	84-99	91
EDTA plasma (n=8)	91-103	97
Cell culture media (n=8)	92-104	97

Linearity

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

Target Information

Synonyms	AFP, FETA, HPAFP, Alpha-fetoglobulin, oralpha fetal protein
Research Area	Cancer, Cardiovascular, Developmental biology

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

BIOMATERIALS (2022) IF: 15.304
Immunology and bioinformatics analysis of injectable organic/inorganic microfluidic microspheres for promoting bone repair

DOI: 10.1016/j.biomaterials.2022.121685

PMID: 35953327
Sample: BMSCs
American Journal of Cancer Research (2021) IF: 6.166
Adamantane-linked isothiourea derivatives suppress the growth of experimental hepatocellular carcinoma via inhibition of TLR4-MyD88-NF-κB signaling

PMID: 33575076
Sample: Tissue homogenate(liver tissue),Serum
Frontiers in Pharmacology (2023) IF: 5.988
Antiangiogenic drugs in combination with seaweed fucoidan: A mechanistic in vitro and in vivo study exploring the VEGF receptor and its downstream signaling molecules in hepatic cancer.

DOI: 10.3389/fphar.2023.1108992

Sample: serum,Liver tissue
JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY (2022) IF: 3.652
Suppressive effects of thymoquinone on the initiation stage of diethylnitrosamine hepatocarcinogenesis in rats

DOI: 10.1002/jbt.23078

PMID: 35437842
Sample: serum
BMC Complementary and Alternative Medicine (2021) IF: 3.659
Oral administration of tartrazine (E102) accelerates the incidence and the development of 7,12-dimethylbenz(a) anthracene (DMBA)-induced breast cancer in rats

DOI: 10.1186/s12906-021-03490-0

PMID: 34972512
Sample: serum
DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY (2017) IF: 2.295
Exploiting bilosomes for delivering bioactive polysaccharide isolated from Enteromorpha intestinalis for hacking hepatocellular carcinoma

DOI: 10.1080/03639045.2017.1402922

Sample: Serum
Evidence-based Complementary and Alternative Medicine (2022) IF: 2.63
Polysaccharide-Rich Fractions from Ganoderma resinaceum (Ganodermataceae) as Chemopreventive Agents in N-Diethylnitrosamine-Induced Hepatocellular Carcinoma in Wistar Rats

DOI: 10.1155/2022/8198859

PMID: 35463072
Sample: liver tissue
DIGESTIVE AND LIVER DISEASE (2016) IF: 2.719
The antitumor activity of a lactosaminated albumin conjugate of doxorubicin in a chemically induced hepatocellular carcinoma rat model compared to sorafenib

DOI: 10.1016/j.dld.2016.10.003

Sample: Serum
NUTRITION AND CANCER-AN INTERNATIONAL JOURNAL (2021) IF: 2.9
Phenolics-Enriched Fraction of Pterospermum Lanceifolium Roxb. efficiently Reverses the Hepatocellular Carcinoma in NDEA-Induced HCC Rats

DOI: 10.1080/01635581.2021.1922716

PMID: 34018459
Sample: Tissue homogenate
Molecular Medicine Reports (2021) IF: 2.952
Kangxianruangan granule‑containing serum mediated inhibition of hepatic oval cell differentiation into hepatocellular carcinoma cells via the Wnt‑1/β‑catenin signaling pathway

DOI: 10.3892/mmr.2021.12571

PMID: 34913065
Sample: WB-F344 cell

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