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AB326009

Human AFP ELISA Kit, Chemiluminescent

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Human AFP ELISA Kit, Chemiluminescent is a 55-min single-wash SimpleStep Ignite™ ELISA for the rapid quantification of Human AFP with higher sensitivity and broader measuring range.

- Single 30 min incubation and single-wash step for results in less than an hour.
- Chemiluminescent detection allows a broad dynamic range: 18.3 - 40000 pg/ml with a sensitivity of 13.9 pg/mL.
- Design your own immunoassay: we also offer the conjugation-ready antibody pair.
- Patent pending

View Alternative Names

HPAFP, AFP, Alpha-fetoprotein, Alpha-1-fetoprotein, Alpha-fetoglobulin

7 Images
Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

SimpleStep ELISA® Ignite technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 55 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

Dynamic range comparison between the Chemiluminescent Human alpha Fetoprotein SimpleStep ELISA, colorimetric Human alpha Fetoprotein SimpleStep ELISA (ab193765), and leading colorimetric competitor. The Chemiluminescent SimpleStep ELISA kit shows increased sensitivity and increased dynamic range.

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

Serum from ten individual healthy human male donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration was determined to be 678.4 pg/mL with a range of 133.9 – 2,257 pg/mL.

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

Interpolated concentrations of native AFP in pregnant human serum and HepG2 cell culture supernatant samples. The concentrations of AFP were measured in duplicates, interpolated from the AFP standard curves and corrected for sample dilution. Undiluted samples are as follows : pregnant human serum 3% and HepG2 cell culture supernatant 1 : 666. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration was determined to be 221.8 ng/mL in neat pregnant human serum and 6,165 ng/mL in neat HepG2 cell culture supernatant.

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

Serum from ten individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration in nine detectable donor samples was determined to be 550.7 pg/mL with a range of 247.9-816.1 pg/mL. Donor #5 was not detectable.

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

Standard curve comparison between the Chemiluminescent Human alpha Fetoprotein SimpleStep ELISA and colorimetric Human alpha Fetoprotein SimpleStep ELISA (ab193765) in Sample Diluent 75BS. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)
  • sELISA

Lab

Sandwich ELISA - Human AFP ELISA Kit, Chemiluminescent (AB326009)

Interpolated concentration of native alpha Fetoprotein was measured in duplicate using the chemiluminescent version of this kit at different sample concentrations. Undiluted samples are as follows : serum 100% and plasma (citrate) 100%. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent 75BS.

Key facts

Detection method

Luminescent

Sample types

Serum, Citrate plasma, EDTA Plasma, Heparin Plasma, Cell culture supernatant, Cell culture media, Urine

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 13.9 pg/mL

Range

18.3 - 40000 pg/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human AFP ELISA Kit, Chemiluminescent (ab326009) is a rapid single-wash sandwich 55-min ELISA to measure Human AFP in Serum, Citrate Plasma, EDTA Plasma, Heparin Plasma, Cell Culture Supernatant, Cell Culture Media, Urine with a sensitivity of 13.9 pg/mL.

How the assay works

Human AFP ELISA Kit, Chemiluminescent (ab326009) SimpleStep Ignite™ ELISA employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colorimetric ELISAs.

Assay Specificity

Our SimpleStep Ignite™ ELISA kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human AFP (ab326009) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature
2. Wash
3. Add ChemiHRP development solution - incubate for 2 min
4. Read the results on a plate reader capable of luminescence detection

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-alpha 1 Fetoprotein antibody [EPR20365-101] - BSA and Azide free (Capture) ab252573
- Anti-alpha 1 Fetoprotein antibody [EPR20364-26] - BSA and Azide free (Detector) ab252632
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Precision

[ { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 2" }, { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 4.5" } ]
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Recovery

[ { "sample": "Serum", "range": "88 - 96 %", "average": "= 91" }, { "sample": "Citrate plasma", "range": "89 - 89 %", "average": "= 89" }, { "sample": "EDTA Plasma", "range": "98 - 102 %", "average": "= 100" }, { "sample": "Heparin Plasma", "range": "94 - 97 %", "average": "= 96" }, { "sample": "Cell culture supernatant", "range": "101 - 113 %", "average": "= 103" }, { "sample": "Cell culture media", "range": "91 - 98 %", "average": "= 94" }, { "sample": "Urine", "range": "116 - 121 %", "average": "= 119" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Product protocols

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Target data

Binds copper, nickel, and fatty acids as well as, and bilirubin less well than, serum albumin. Only a small percentage (less than 2%) of the human AFP shows estrogen-binding properties.
See full target information AFP
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websiteProtocolBooklet
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com