Human CXCL5/ENA-78 Quantikine ELISA Kit

R&D Systems | Catalog # DX000

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (10 µL), Serum (25 µL), EDTA Plasma (50 µL), Platelet-poor Heparin Plasma (50 µL), Citrate Plasma (50 µL)

Sensitivity

15 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor Heparin Plasma, Citrate Plasma)
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Product Summary for Human CXCL5/ENA-78 Quantikine ELISA Kit

The Quantikine Human ENA-78 Immunoassay is a 4.5 hour solid phase ELISA designed to measure human ENA-78 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human ENA-78 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant human ENA-78 accurately. Results obtained using natural human ENA-78 showed dose curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human ENA-78.

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 ENA-78

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not 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) 112 251 1096 129 278 1085
Standard Deviation 6.0 14.6 44.9 12.6 26.8 96.5
CV% 5.4 5.8 4.1 9.8 9.6 8.9

Citrate Plasma, EDTA Plasma, Platelet-poor 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) 113 244 995 109 240 989
Standard Deviation 9.4 12.9 37.8 10.1 17.8 65.8
CV% 8.3 5.3 3.8 9.3 7.4 6.7

Recovery for Human CXCL5/ENA-78 Quantikine ELISA Kit

The recovery of ENA-78 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 101 95-107
Citrate Plasma (n=5) 100 94-106
EDTA Plasma (n=5) 98 92-104
Heparin Plasma (n=5) 98 93-103
Serum (n=5) 101 93-109

Linearity

To assess the linearity of the assay, five samples containing or spiked with high concentrations of ENA-78 in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human CXCL5/ENA-78 ELISA Linearity

Scientific Data Images for Human CXCL5/ENA-78 Quantikine ELISA Kit

Human CXCL5/ENA-78 ELISA Cell Culture Supernate Standard Curve

Human CXCL5/ENA-78 ELISA Cell Culture Supernate Standard Curve

Human CXCL5/ENA-78 ELISA Serum/Plasma Standard Curve

Human CXCL5/ENA-78 ELISA Serum/Plasma 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: CXCL5/ENA-78

CXCL5/Epithelial Cell-derived Neutrophil-activating Peptide (ENA-78), is a member of the CXC subfamily of chemokines. Full-length CXCL5/ENA-78 is 114 amino acids (aa) in length with a predicted molecular weight of 12 kDa. Following the removal of the signal peptide, bioactive CXCL5/ENA-78 is 78 aa in length. CXCL5/ENA-78 can be N-terminally cleaved by Cathepsin G and Chymotrypsin to CXCL5/ENA-74 (74 aa) and CXCL5/ENA-70 (70 aa), which show increased potency relative to CXCL5/ENA-78. While murine LIX was thought to be an ortholog to human CXCL5/ENA-78, genome-wide analysis and a consensus in the field suggests that human CXCL5/ENA-78 does not have a true murine ortholog. CXCL5/ENA-78 is upregulated at sites of inflammation and is expressed by multiple hematopoietic cell types, fibroblasts, endothelial cells, vascular smooth muscle cells, and adipocytes. It acts as a chemoattractant for neutrophils, has angiogenic properties, and contributes to cancer progression.

Alternate Names

CXCL5, ENA-78, ENA78

Entrez Gene IDs

6374 (Human)

Gene Symbol

CXCL5

Additional CXCL5/ENA-78 Products

Product Documents for Human CXCL5/ENA-78 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 CXCL5/ENA-78 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 CXCL5/ENA-78 Quantikine ELISA Kit

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  • Human CXCL5/ENA-78 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Media
    Verified Customer | Posted 09/07/2017
    Curve fit in Prism and interpolated the sample concentrations. Kit worked well.
    Human CXCL5/ENA-78 Quantikine ELISA Kit DX000

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Protocols

View specific protocols for Human CXCL5/ENA-78 Quantikine ELISA Kit (DX000):

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. 200 µL Assay Diluent
  3.   Add 200 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 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 twice for a total of 3 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 3 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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