alpha-Fetoprotein (AFP), is albuminoid superfamily protein that is synthesized in the fetus primarily by the liver, yolk sac, and tissues of gastrointestinal origin. It is one of the earliest markers of the hepatocyte lineage. AFP acts as a carrier protein for steroids, bilirubin, fatty acids, retinoids, and flavonoids. In addition, it can exert immunosuppressive activity, regulate cell proliferation and apoptosis, initiate intracellular signaling, and contribute to cell invasion. Altered levels of both fetal and maternal AFP have been associated with hypothyroidism, autoimmune disorders, and heart defects. Low maternal serum AFP levels are associated with a higher incidence of Down syndrome, whereas higher levels are associated with spina bifida and anencephaly. Elevated AFP levels are also coincident with liver, stomach, and germ cell cancers.
Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
R&D Systems | Catalog # MAFP00
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)
Sensitivity
0.029 ng/mL
Assay Range
0.156-10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
The Quantikine Mouse alpha -Fetoprotein/AFP Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse AFP levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse AFP and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse AFP accurately. Results obtained using natural mouse AFP showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse AFP.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Mouse
Specificity
Natural and recombinant mouse AFP.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 0.404 | 1.12 | 3.30 | 0.457 | 1.20 | 3.38 |
| Standard Deviation | 0.013 | 0.026 | 0.091 | 0.031 | 0.066 | 0.158 |
| CV% | 3.2 | 2.3 | 2.8 | 6.8 | 5.5 | 4.7 |
Recovery for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
The recovery of mouse AFP spiked to levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 98 | 93-105 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse AFP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data Images for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
Mouse alpha-Fetoprotein/AFP ELISA Standard Curve
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: alpha-Fetoprotein/AFP
Additional alpha-Fetoprotein/AFP Products
Product Documents for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
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.
Product Specific Notices for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit
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5 Customer Ratings
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Sample Tested: Plasmas from mice fed with HFDVerified Customer | Posted 01/08/2022I am very satisfied by this product.
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Sample Tested: SerumVerified Customer | Posted 08/06/2018
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Sample Tested: SerumVerified Customer | Posted 03/23/2018Easy to use with reliable results. Would recommend.
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Sample Tested: mouse serumVerified Customer | Posted 03/23/2018
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Sample Tested: Mouse sera diluted 120x in Calibrator Diluent RD5-26 (diluted 1:4)Verified Customer | Posted 01/13/2017
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Protocols
View specific protocols for Mouse alpha-Fetoprotein/AFP Quantikine ELISA Kit (MAFP00):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
