Mouse CCL12/MCP-5 Quantikine ELISA Kit

(4 citations)
(1 Review)
  
  • 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 (25 uL), EDTA Plasma (25 uL)
  • Sensitivity
    2.1 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse MCP-5
  • 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.
Product Summary
The Quantikine Mouse MCP-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MCP-5 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse MCP-5 and antibodies raised against the recombinant factor. Results obtained for naturally occurring mouse MCP-5 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 mouse MCP-5.

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, Serum, EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean32853183186318
Standard Deviation1.83.5182.87.933
CV%5.64.15.799.210.4

Recovery

The recovery of mouse MCP-5 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 94 81-119
EDTA Plasma (n=6) 99 84-114
Serum (n=6) 93 80-108
Linearity
To assess the linearity of the assay, six samples containing and/or spiked with high concentrations of mouse MCP-5 in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse CCL12/MCP-5 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CCL12/MCP-5
Mouse MCP-5, also known as CCL12, encodes a 104 amino acid (aa) precursor protein with a 22 aa hydrophobic signal sequence that is cleaved to generate a 82 aa residue mature protein. MCP-5 is expressed constitutively in the thymus and lymph nodes. Under inflammatory conditions, MCP-5 expression is also induced in activated macrophages and mast cells.
    • Entrez Gene IDs
      20293 (Mouse);
    • Alternate Names
      C-C motif chemokine 12; CCL12; chemokine (C-C motif) ligand 12; MCP5; MCP-5; Scya12;
    Related Research Areas
    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. 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. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    4 Citations: Showing 1 - 4
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    Species
    Sample Type
    1. Varying susceptibility of pulmonary cytokine production to lipopolysaccharide in mice.
      Authors: Alm AS, Li K, Yang D, Andersson R, Lu Y, Wang X
      Cytokine, 2010;49(3):256-63.
      Species: Mouse
      Sample Type: BALF
    2. Src homology 2 domain-containing inositol 5-phosphatase 1 deficiency leads to a spontaneous allergic inflammation in the murine lung.
      Authors: Oh SY, Zheng T, Bailey ML, Barber DL, Schroeder JT, Kim YK, Zhu Z
      J. Allergy Clin. Immunol., 2007;119(1):123-31.
      Species: Mouse
      Sample Type: BALF
    3. Type I IFN modulates host defense and late hyperinflammation in septic peritonitis.
      Authors: Weighardt H, Kaiser-Moore S, Schlautkotter S, Rossmann-Bloeck T, Schleicher U, Bogdan C, Holzmann B
      J. Immunol., 2006;177(8):5623-30.
      Species: Mouse
      Sample Type: Peritoneal Lavage
    4. Nuclear factor-kappaB affects tumor progression in a mouse model of malignant pleural effusion.
      Authors: Stathopoulos GT, Zhu Z, Everhart MB, Kalomenidis I, Lawson WE, Bilaceroglu S, Peterson TE, Mitchell D, Yull FE, Light RW, Blackwell TS
      Am. J. Respir. Cell Mol. Biol., 2005;34(2):142-50.
      Species: Mouse
      Sample Type: Serum
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    Average Rating: 5 (Based on 1 review)

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     Mouse CCL12/MCP-5 Quantikine ELISA Kit MCC120
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      Anonymous 05/19/2017
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     Mouse CCL12/MCP-5 Quantikine ELISA Kit MCC120
    : Mouse CCL12/MCP-5 Quantikine ELISA Kit [MCC120]

    Summary

    Sample Testedmouse lung tissue

    Other Experimental Details

    Other Experimental DetailsThe ELISA worked absolutely fine in Mouse lung tissue homogenate made in RIPA buffer.1:4 dilution works absolutely fine with this kit if 15-25 mg of lung tissue is digested in 250ul of RIPA buffer. To make homogenates other buffer like PBS can also be used. Thanks

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