Mouse LIX Quantikine ELISA Kit

  (4 citations)     
Datasheet / CoA / SDS
Product Details
Assay Procedure
Citations (4)
FAQs
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  • 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), Tissue Homogenates (50 uL), Serum (10 uL), Platelet-poor EDTA Plasma (13 uL), Platelet-poor Heparin Plasma (13 uL), Urine (13 uL)
  • Sensitivity
    1.74 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Tissue Homogenates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant mouse LIX
  • 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 Mouse CXCL5/LIX Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse LIX levels in cell culture supernates, tissue lysates, serum, platelet-poor plasma, and urine. It contains E. coli-expressed recombinant mouse LIX and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse LIX accurately. Results obtained using natural mouse LIX showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse LIX.

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, Tissue Homogenates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 42.7 133 407 41.8 120 393
Standard Deviation 1.72 5.93 10.9 2.81 4.64 20.3
CV% 4 4.5 2.7 6.7 3.9 5.2

Recovery

The recovery of mouse LIX spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 114 105-120
Platelet-poor EDTA Plasma (n=4) 100 92-109
Platelet-poor Heparin Plasma (n=4) 91 85-99
Serum (n=4) 87 82-94
Tissue Homogenates (n=4) 100 87-113
Urine (n=4) 91 84-96
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse LIX were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
 LIX [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: LIX
LPS-induced CXC chemokine (LIX) is a member of the murine chemokine family. It is 132 amino acids (aa) in length with a predicted molecular weight of 14 kDa prior to cleavage of the N-terminal signal peptide (aa 1–40). LIX has no true human ortholog but is closely related to human CXCL5/ENA-78 and CXCL6/GCP-2. It is widely expressed, and is secreted by multiple cell types including fibroblasts, thymic epithelium, platelets, vascular endothelium, and hepatocytes. LIX acts as a proinflammatory chemokine and is chemotactic for neutrophils.
  • Entrez Gene IDs:
    20311 (Mouse); 60665 (Rat)
  • Alternate Names:
    LIX
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. Upregulation of SOCS3 in lung CD4+ T cells in a mouse model of chronic PA lung infection and suppression of Th17?mediated neutrophil recruitment in exogenous SOCS3 transfer in vitro
    Authors: FM Ding, RM Liao, YQ Chen, GG Xie, PY Zhang, P Shao, M Zhang
    Mol Med Rep, 2017;0(0):.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  2. Sitagliptin, a dipeptidyl peptidase-4 inhibitor, suppresses CXCL5 and SDF-1 and does not accelerate intestinal neoplasia formation in Apc(Min/+) mice fed a high-fat diet
    Authors: K Fujiwara, T Inoue, Y Henmi, Y Hirata, Y Naka, A Hara, K Kakimoto, S Nouda, T Okada, K Kawakami, T Takeuchi, K Higuchi
    Oncol Lett, 2017;14(4):4355-4360.
    Species: Mouse
    Sample Type: Plasma
  3. Sharpin Controls Osteogenic Differentiation of Mesenchymal Bone Marrow Cells.
    Authors: Jeschke A, Catala-Lehnen P, Sieber S, Bickert T, Schweizer M, Koehne T, Wintges K, Marshall R, Mautner A, Duchstein L, Otto B, Horst A, Amling M, Kreienkamp H, Schinke T
    J Immunol, 2015;195(8):3675-84.
    Species: Mouse
    Sample Type: Serum
  4. Decreased plasma cytokines are associated with low platelet counts in aplastic anemia and immune thrombocytopenic purpura.
    J. Thromb. Haemost., 2012;10(8):1616-23.
    Species: Mouse
    Sample Type: Plasma

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