Human CCL20/MIP-3 alpha DuoSet ELISA

R&D Systems | Catalog # DY360

R&D Systems
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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

15.6-1000 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 CCL20/MIP-3 alpha 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
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Product Summary for Human CCL20/MIP-3 alpha DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CCL20/MIP-3alpha. 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

Label

HRP

Scientific Data Images for Human CCL20/MIP-3 alpha DuoSet ELISA

Human CCL20 / MIP-3 alpha ELISA Standard Curve

Human CCL20 / MIP-3 alpha ELISA Standard Curve

Kit Contents for Human CCL20/MIP-3 alpha 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 # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

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 equivalent

Reagent Diluent*

Blocking Buffer*

Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)

Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)

Microplates: (Catalog # DY990), or equivalent

Plate Sealers: (Catalog # DY992), or equivalent

*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.

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: CCL20/MIP-3 alpha

CCL20, also known as MIP-3 alpha, is a chemokine that plays an important role in dendritic cell trafficking and recruitment and activation of T cells. It is upregulated in monocytes, T cells, endothelial cells, epithelial cells, and fibroblasts following inflammatory stimulation. CCL20 signals through CCR6 which is expressed on memory T cells, B cells and bone marrow-derived dendritic cells.

Alternate Names

exodus-1, LARC, MIP-3 alpha, MIP3 alpha

Entrez Gene IDs

6364 (Human); 20297 (Mouse); 29538 (Rat)

Gene Symbol

CCL20

Additional CCL20/MIP-3 alpha Products

Product Documents for Human CCL20/MIP-3 alpha 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.

Product Specific Notices for Human CCL20/MIP-3 alpha DuoSet ELISA

For research use only

Citations for Human CCL20/MIP-3 alpha DuoSet ELISA

Customer Reviews for Human CCL20/MIP-3 alpha DuoSet ELISA (4)

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Showing  1 - 4 of 4 reviews Showing All
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  • CCL20 in HCT116 cell line
    Name: Anonymous
    Sample Tested: HCT-116 human colorectal carcinoma cell line
    Species: Human
    Verified Customer | Posted 02/03/2026
    Supernatant of HCT116 cells ELISA assay for CCL20
    Dilution factor of 1.33
    Human CCL20/MIP-3 alpha DuoSet ELISA DY360
  • CCL20 in supernatant of MDA-231 cells
    Name: Caterina Suelzu
    Sample Tested: MDA-MB-231 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 02/03/2026
    CCL20 levels in MDA-231 cells
    Samples of media were diluted three times.
    Human CCL20/MIP-3 alpha DuoSet ELISA DY360
  • Human CCL20/MIP-3 alpha DuoSet ELISA
    Name: Laboratorio Inmuno-Oncología
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 11/04/2020
    Human CCL20/MIP-3 alpha DuoSet ELISA DY360
  • Human CCL20/MIP-3 alpha DuoSet ELISA
    Name: Anonymous
    Sample Tested: Purified Standard
    Verified Customer | Posted 08/23/2016
    Human CCL20/MIP-3 alpha DuoSet ELISA DY360

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Showing  1 - 4 of 4 reviews Showing All

Protocols

View specific protocols for Human CCL20/MIP-3 alpha DuoSet ELISA (DY360):

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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