Human FAP DuoSet ELISA

Catalog # Availability Size / Price Qty
Ancillary Products Available
Human Fibroblast Activation Protein alpha / FAP ELISA Standard Curve
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Product Details
Citations (9)
Supplemental Products

Human FAP DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Fibroblast Activation Proteinalpha/FAP. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human Fibroblast Activation Protein alpha / FAP ELISA Standard Curve

Product Datasheets

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Preparation and Storage

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: Fibroblast Activation Protein alpha/FAP

FAP (fibroblast activation protein-alpha; also known as seprase) is a type II transmembrane serine protease that degrades gelatin, Type I collagen, and N-terminal prolines. FAP is induced on reactive stromal fibroblasts in epithelial cancers, sarcomas, and granulation tissue, and may play roles in tumor invasion, tissue remodeling, and wound repair.

Entrez Gene IDs:
2191 (Human); 14089 (Mouse); 102134935 (Cynomolgus Monkey)
Alternate Names:
170 kDa melanoma membrane-bound gelatinase; DKFZp686G13158; DPPIV; EC 3.4.21.-; FAP; FAPA; Fibroblast Activation Protein alpha; fibroblast activation protein, alpha; Integral membrane serine protease; Seprase; vibronectin

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human FAP DuoSet ELISA

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. Effect of &alpha2-plasmin inhibitor heterogeneity on the risk of venous thromboembolism
    Authors: B Baráth, R Bogáti, T Miklós, J Kállai, ZA Mezei, Z Bereczky, L Muszbek, É Katona
    Thrombosis Research, 2021;203(0):110-116.
    Species: Human
    Sample Types: Plasma
  2. Incorporation of &alpha2-Plasmin Inhibitor into Fibrin Clots and Its Association with the Clinical Outcome of Acute Ischemic Stroke Patients
    Authors: Z Bagoly, B Baráth, R Orbán-Kálm, I Szegedi, R Bogáti, F Sarkady, L Csiba, É Katona
    Biomolecules, 2021;11(3):.
    Species: Human
    Sample Types: Plasma
  3. Colorectal cancer cell intrinsic fibroblast activation protein alpha binds to Enolase1 and activates NF-&kappaB pathway to promote metastasis
    Authors: Z Yuan, H Hu, Y Zhu, W Zhang, Q Fang, T Qiao, T Ma, M Wang, R Huang, Q Tang, F Gao, C Zou, X Gao, G Wang, X Wang
    Cell Death & Disease, 2021;12(6):543.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Regulation of Fibroblast Activation Protein by Transforming Growth Factor Beta-1 in Glioblastoma Microenvironment
    Authors: E Krepela, Z Vanickova, P Hrabal, M Zubal, B Chmielova, E Balaziova, P Vymola, I Matrasova, P Busek, A Sedo
    International Journal of Molecular Sciences, 2021;22(3):.
  5. Identification of the transcription factor Miz1 as an essential regulator of diphthamide biosynthesis using a CRISPR-mediated genome-wide screen
    Authors: J Liu, Z Zuo, M Zou, T Finkel, S Liu
    PLoS Genet, 2020;16(10):e1009068.
    Species: Human
    Sample Types: Cell Fragments
  6. Altered Tissue and Plasma Levels of Fibroblast Activation Protein-&alpha (FAP) in Renal Tumours
    Authors: JD Solano-Itu, P Errarte, MC Etxezarrag, E Echevarria, J Angulo, JI López, G Larrinaga
    Cancers (Basel), 2020;12(11):.
    Species: Human
    Sample Types: Plasma
  7. Selective Homogeneous Assay for Circulating Endopeptidase Fibroblast Activation Protein (FAP)
    Authors: TW Bainbridge, DR Dunshee, NM Kljavin, NJ Skelton, J Sonoda, JA Ernst
    Sci Rep, 2017;7(1):12524.
    Species: Human
    Sample Types: Serum
  8. Circulating fibroblast activation protein activity and antigen levels correlate strongly when measured in liver disease and coronary heart disease
    Authors: S Uitte de W, FM Keane, DG Bowen, JJMC Malfliet, HE Zhang, B Maneck, GW McCaughan, FWG Leebeek, DC Rijken, MD Gorrell
    PLoS ONE, 2017;12(6):e0178987.
    Species: Human
    Sample Types: Plasma
  9. Heterogeneity of molecular forms of dipeptidyl peptidase-IV and fibroblast activation protein in human glioblastomas.
    Authors: Matrasova I, Busek P, Balaziova E, Sedo A
    Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub, 0;161(3):252-260.
    Species: Human
    Sample Types: Tissue Homogenates


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