Human CCL20/MIP-3 alpha Quantikine ELISA Kit

R&D Systems | Catalog # DM3A00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL)

Sensitivity

0.87 pg/mL

Assay Range

7.8-500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human CCL20/MIP-3 alpha Quantikine ELISA Kit

The Quantikine Human MIP-3 alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure MIP-3 alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-3 alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring human MIP-3 alpha showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural human MIP-3 alpha.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human MIP-3 alpha

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

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 (pg/mL) 66.2 180 372 68.7 186 357
Standard Deviation 1.84 3.83 15.1 4.51 13.0 28.8
CV% 2.8 2.1 4.1 6.6 6.9 8.1

EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 55.9 161 323 72.5 200 386
Standard Deviation 1.62 4.28 9.10 6.05 17.9 35.3
CV% 2.9 2.7 2.8 8.3 8.9 9.1

Recovery for Human CCL20/MIP-3 alpha Quantikine ELISA Kit

The recovery of MIP-3 alpha spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=8) 101 88-115
EDTA Plasma (n=5) 104 97-115
Heparin Plasma (n=5) 104 98-112
Serum (n=5) 103 94-112

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of MIP-3 alpha in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human CCL20/MIP-3 alpha ELISA Linearity

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

Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6U Standard Curve

Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6U Standard Curve

Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6-21 Standard Curve

Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6-21 Standard Curve

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 Quantikine ELISA Kit

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 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human CCL20/MIP-3 alpha Quantikine ELISA Kit

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  • Human CCL20/MIP-3 alpha Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 01/20/2017
    This kit is a very nice tool in determining the level of MIP-3alpha in the cell culture spent media.
    Human CCL20/MIP-3 alpha Quantikine ELISA Kit DM3A00
  • Human CCL20/MIP-3 alpha Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Stimulated Human Bronchial Epithelial Cell Line (BEAS-2B)
    Verified Customer | Posted 01/03/2017
    This kit is an excellent tool in determining the level of MIP-3a in the culture media.

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Protocols

View specific protocols for Human CCL20/MIP-3 alpha Quantikine ELISA Kit (DM3A00):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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