Canine CCL2/MCP-1 Quantikine ELISA Kit

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CACP00
Canine CCL2/JE/MCP-1 ELISA Standard Curve
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Citations (5)
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Canine CCL2/MCP-1 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
10.8 pg/mL
Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant canine MCP-1
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.

Product Summary

The Quantikine Canine MCP-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine MCP-1 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine MCP-1 and antibodies raised against the recombinant protein. Results obtained for naturally occurring canine MCP-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural canine MCP-1.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times 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, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 43 43 43
Mean (pg/mL) 37.1 144 632 42.7 133 655
Standard Deviation 2.2 5.8 22 5.4 7.5 30
CV% 5.9 4 3.5 12.6 5.6 4.6

Recovery

The recovery of canine MCP-1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 112 106-121
EDTA Plasma (n=4) 110 102-116
Heparin Plasma (n=4) 103 96-112
Serum (n=4) 93 84-102

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of canine MCP-1 in each matrix were diluted with Calibrator Diluent and assayed.
Canine CCL2/JE/MCP-1 ELISA Linearity

Scientific Data

Canine CCL2/JE/MCP-1 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL2/JE/MCP-1

CCL2/JE/MCP-1, is a chemokine that binds the receptor CCR2 and induces the chemoattraction of mononuclear cells. It induces the activation of monocytes, NK cells, lymphocytes, and basophils. Additionally, CCL2 promotes Th2 polarization in CD4+ T cells, and CCL2-mediated recruitment of monocytes to sites of inflammation contributes to disease severity in atherosclerosis, multiple sclerosis, and allergic asthma. Endogenous proteolytic trimming of CCL2 at the N-terminus, including the N-terminal pyrrolidone carboxylic acid-modified glutamine, downregulates activity but not receptor binding.

Entrez Gene IDs:
6347 (Human); 20296 (Mouse); 24770 (Rat); 403981 (Canine)
Alternate Names:
C-C motif chemokine ligand 2; CCL2; GDCF-2; HC11; HSMCR30; MCAF; Mcp1; MCP-1; SCYA2; SMC-CF
&#9888; 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.

Assay Procedure

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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Tap the plate gently for 1 minute. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Canine CCL2/MCP-1 Quantikine ELISA Kit

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.

5 Citations: Showing 1 - 5
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  1. Serum and urinary monocyte chemoattractant protein-1 as markers of inflammation and renal damage in dogs with naturally occurring leishmaniosis
    Authors: Pantaleo, V;Furlanello, T;Ventura, L;Solano-Gallego, L;
    Parasites & vectors
    Species: Canine
    Sample Types: Serum, Urine
  2. Ability of different assay platforms to measure renal biomarker concentrations during ischaemia-reperfusion acute kidney injury in dogs
    Authors: J Davis, G Rossi, DW Miller, RE Cianciolo, AL Raisis
    Res Vet Sci, 2020-11-12;0(0):.
    Species: Canine
    Sample Types: Plasma
  3. Up-regulated inflammatory signatures of the spinal cord in canine degenerative myelopathy
    Authors: K Hashimoto, Y Kobatake, R Asahina, O Yamato, MS Islam, H Sakai, H Nishida, S Maeda, H Kamishina
    Res Vet Sci, 2020-11-05;0(0):.
    Species: Canine
    Sample Types: Whole Tissue
    Applications: IHC
  4. Evaluation of potential serum biomarkers of hepatic fibrosis and necroinflammatory activity in dogs with liver disease
    Authors: C Raghu, J Ekena, JM Cullen, CB Webb, LA Trepanier
    J. Vet. Intern. Med., 2018-02-27;0(0):.
    Species: Canine
    Sample Types: Serum
  5. CC chemokine ligand 2 and CXC chemokine ligand 8 as neutrophil chemoattractant factors in canine idiopathic polyarthritis
    Authors: Naoaki Matsuki
    Vet. Immunol. Immunopathol., 2016-09-23;182(0):52-58.
    Species: Canine
    Sample Types: Synovial Fluid

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