Human FAP DuoSet ELISA

R&D Systems | Catalog # DY3715

R&D Systems
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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

62.5-4000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human FAP DuoSet ELISA 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
View all Fibroblast Activation Protein alpha/FAP ELISA Kits »

Product Summary for Human FAP DuoSet ELISA

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 Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human FAP DuoSet ELISA

Human Fibroblast Activation Protein alpha / FAP ELISA Standard Curve

Human Fibroblast Activation Protein alpha / FAP ELISA Standard Curve

Kit Contents for Human FAP DuoSet ELISA

  • 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)

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: Fibroblast Activation Protein alpha/FAP

FAP (also known as seprase) is a transmembrane serine protease, and a soluble and enzymatically active form of FAP known as antiplasmin-cleaving enzyme (APCE) circulates in human plasma. FAP is expressed on reactive stromal fibroblasts in tumor tissue and wound healing and on synoviocytes in rheumatoid arthritis. It exhibits dipeptidyl peptidase activity with substrate specificity similar to DPPIV/CD26, which is specific for N-terminal Xaa-Pro sequences. FAP is also an endopeptidase that can degrade Gelatin, Collagens I and IV, Fibronectin, and Laminin as well as several peptide hormones (e.g. Neuropeptide Y, Brain Natriuretic Peptide, Substance P, Peptide YY, and Incretins). The enzymatic activity is dependent on FAP association with DPPIV on the cell surface. The matrix-dedgrading activity of FAP contributes to tumor cell migration and invasion. In addition, FAP can enhance tumor cell growth by limiting the development of anti-tumor immunity.

Alternate Names

FAP, Seprase

Entrez Gene IDs

2191 (Human); 14089 (Mouse); 102134935 (Cynomolgus Monkey)

Gene Symbol

FAP

Additional Fibroblast Activation Protein alpha/FAP Products

Product Documents for Human FAP DuoSet ELISA

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.

Download SDS

Product Specific Notices for Human FAP DuoSet ELISA

For research use only

Citations for Human FAP DuoSet ELISA

Customer Reviews for Human FAP DuoSet ELISA (2)

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  • Human FAP DuoSet ELISA
    Name: Sergi Marco
    Sample Tested: Plasma
    Verified Customer | Posted 05/22/2023
    Human Plasma samples diluted between 1/100 and 1/200 fit perfectly in the standard curve.
    Human FAP DuoSet ELISA DY3715
  • Human FAP DuoSet ELISA
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 09/23/2019
    we used this kit for quantifying human FAP in serum samples. This kit works well and detects FAP very efficiently in human serum.
    Human FAP DuoSet ELISA DY3715

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Protocols

View specific protocols for Human FAP DuoSet ELISA (DY3715):

GENERAL ELISA PROTOCOL

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.

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