Human alpha-Fetoprotein Quantikine ELISA Kit

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DAFP00
Control Products Available
Human alpha-Fetoprotein/AFP ELISA Calibrator Diluent RD5-24 Standard Curve
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Product Details
Procedure
Citations (9)
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Human alpha-Fetoprotein Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
0.046 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural human AFP
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
Interference
Interference observed with other substances.

Product Summary

The Quantikine Human AFP immunoassay is a 4.5 hour solid phase ELISA designed to measure human AFP in cell culture supernates, serum, and plasma.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 2.89 6.02 12.3 2.65 5.62 11.7
Standard Deviation 0.109 0.219 0.441 0.158 0.261 0.549
CV% 3.8 3.6 3.6 6 4.6 4.7

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 3.15 6.62 13.2 3.22 6.79 14.1
Standard Deviation 0.08 0.149 0.447 0.215 0.382 0.93
CV% 2.5 2.3 3.4 6.7 5.6 6.6

Recovery

The recovery of AFP spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 95-103
EDTA Plasma (n=4) 104 94-114
Heparin Plasma (n=4) 103 95-115
Serum (n=4) 103 86-115

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of AFP were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human alpha-Fetoprotein/AFP ELISA Linearity

Data Examples

Human alpha-Fetoprotein/AFP ELISA Calibrator Diluent RD5-24 Standard Curve

Human alpha-Fetoprotein/AFP ELISA Calibrator Diluent RD5-33 Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: alpha-Fetoprotein/AFP

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.

Entrez Gene IDs:
174 (Human); 11576 (Mouse); 24177 (Rat)
Alternate Names:
AFP; Alpha-1-fetoprotein; alpha-fetoglobulin; alpha-Fetoprotein; DSCAM2; FETA; HP; HPAFP
⚠ 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.

Assay Procedure

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human alpha-Fetoprotein Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

9 Citations: Showing 1 - 9
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  1. Maternal biomarkers for fetal heart failure in fetuses with congenital heart defects or arrhythmias
    Authors: T Miyoshi, H Hosoda, M Nakai, K Nishimura, M Miyazato, K Kangawa, T Ikeda, J Yoshimatsu, N Minamino
    Am. J. Obstet. Gynecol., 2019;220(1):104.e1-104.e1.
    Species: Human
    Sample Types: Serum
  2. Identification of anti-SF3B1 autoantibody as a diagnostic marker in patients with hepatocellular carcinoma
    Authors: HM Hwang, CK Heo, HJ Lee, SS Kwak, WH Lim, JS Yoo, DY Yu, KJ Lim, JY Kim, EW Cho
    J Transl Med, 2018;16(1):177.
    Species: Human
    Sample Types: Serum
  3. Serum long noncoding RNA LRB1 as a potential biomarker for predicting the diagnosis and prognosis of human hepatocellular carcinoma
    Authors: ZF Wang, R Hu, JM Pang, GZ Zhang, W Yan, ZN Li
    Oncol Lett, 2018;16(2):1593-1601.
    Species: Human
    Sample Types: Serum
  4. Knockdown of beta-catenin with dicer-substrate siRNAs reduces liver tumor burden in vivo.
    Authors: Dudek H, Wong D, Arvan R, Shah A, Wortham K, Ying B, Diwanji R, Zhou W, Holmes B, Yang H, Cyr W, Zhou Y, Shah A, Farkiwala R, Lee M, Li Y, Rettig G, Collingwood M, Basu S, Behlke M, Brown B
    Mol Ther, 2014;22(1):92-101.
    Species: Human
    Sample Types: Serum
  5. Proteomics mapping of cord blood identifies haptoglobin "switch-on" pattern as biomarker of early-onset neonatal sepsis in preterm newborns.
    Authors: Buhimschi CS, Bhandari V, Dulay AT, Nayeri UA, Abdel-Razeq SS, Pettker CM, Thung S, Zhao G, Han YW, Bizzarro M, Buhimschi IA
    PLoS ONE, 2011;6(10):e26111.
    Species: Human
    Sample Types: Serum
  6. DLK1, a serum marker for hepatoblastoma in young infants.
    Authors: Falix FA, Aronson DC, Lamers WH, Hiralall JK, Seppen J
    Pediatr Blood Cancer, 2011;59(0):743.
    Species: Human
    Sample Types: Serum
  7. A hypoxia- and alpha-fetoprotein-dependent oncolytic adenovirus exhibits specific killing of hepatocellular carcinomas.
    Authors: Kwon OJ, Kim PH, Huyn S, Wu L, Kim M, Yun CO
    Clin. Cancer Res., 2010;16(24):6071-82.
    Species: Human
    Sample Types: Serum
  8. Identifying serological biomarkers of hepatocellular carcinoma using surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy.
    Authors: Wu FX, Wang Q, Zhang ZM, Huang S, Yuan WP, Liu JY, Ban KC, Zhao YN
    Cancer Lett., 2009;279(2):163-70.
    Species: Human
    Sample Types: Serum
  9. A novel approach and protocol for discovering extremely low-abundance proteins in serum.
    Authors: Tanaka Y, Akiyama H, Kuroda T, Jung G, Tanahashi K, Sugaya H, Utsumi J, Kawasaki H, Hirano H
    Proteomics, 2006;6(17):4845-55.
    Species: Human
    Sample Types: Serum

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