Rat CXCL2/CINC-3 Quantikine ELISA Kit

(13 citations)   
  • 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
    2.7 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant rat CINC-3
  • 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 Rat CINC-3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure rat CINC-3 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat CINC-3 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural rat CINC-3 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 naturally occurring rat CINC-3.

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, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020504650
Mean1113451012105308877
Standard Deviation3.911.939.110.128.573.2
CV%3.53.43.99.69.38.3

Recovery

The recovery of rat CINC-3 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) 97 86-107
EDTA Plasma (n=6) 92 86-99
Heparin Plasma (n=6) 99 85-105
Serum (n=6) 97 83-106
Linearity
To assess the linearity of the assay, samples containing rat CINC-3 in each matrix were diluted with Calibrator Diluent and then assayed.
Rat CXCL2/CINC-3 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CXCL2/GRO beta/MIP-2/CINC-3
CXCL2/GRO beta, also called MIP-2 in mouse and CINC-3 in rat, is a member of the CXC chemokine family. Human CXCL2/GRO beta is 107 amino acids (aa) in length with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 70% and 71% aa sequence identity with the human protein, respectively. N-terminal aa 1-4 of CXCL2/GRO beta can be post-translationally cleaved which confers enhanced hematopoietic bioactivity. CXCL2/GRO beta is produced by a variety of cell types including monocytes and macrophages at sites of inflammation and is chemotactic for granulocytes, including neutrophils.
    • Entrez Gene IDs
      2920 (Human); 20310 (Mouse); 114105 (Rat);
    • Alternate Names
      chemokine (C-X-C motif) ligand 2; CINC-2a; CINC3; CINC-3; C-X-C Motif Chemokine 2; CXCL2; GRO beta; GRO2 oncogene; GRO2; GROB; Gro-beta; Growth-regulated protein beta; Macrophage inflammatory protein 2-alpha; melanoma growth stimulatory activity beta; MGSA beta; MGSA-b; MGSA-beta; MIP2; MIP-2; MIP2A; MIP-2a; MIP2-alpha; SCYB2;
    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.

    13 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Assessment of Pulmonary Toxicity Induced by Inhaled Toner with External Additives
      Authors: T Tomonaga, H Izumi, Y Yoshiura, T Myojo, T Oyabu, BW Lee, T Okada, Y Li, K Kawai, T Higashi, Y Morimoto
      Biomed Res Int, 2017;2017(0):4245309.
      Species: Mouse
      Sample Type: Tissue Homogenates
    2. NF-kappaB and STAT1 control CXCL1 and CXCL2 gene transcription.
      Authors: Burke S, Lu D, Sparer T, Masi T, Goff M, Karlstad M, Collier J
      Am J Physiol Endocrinol Metab, 2014;306(2):E131-49.
      Species: Rat
      Sample Type: Cell Culture Supernates
    3. A short course of infusion of a hydrogen sulfide-donor attenuates endotoxemia induced organ injury via stimulation of anti-inflammatory pathways, with no additional protection from prolonged infusion.
      Cytokine, 2013;61(2):614-21.
      Species: Rat
      Sample Type: BALF
    4. Mechanical ventilation enhances HMGB1 expression in an LPS-induced lung injury model.
      Authors: Ding N, Wang F, Xiao H, Xu L, She S
      PLoS ONE, 2013;8(9):e74633.
      Species: Rat
      Sample Type: BALF
    5. P2X7 receptor activation induces CXCL2 production in microglia through NFAT and PKC/MAPK pathways.
      Authors: Shiratori M, Tozaki-Saitoh H, Yoshitake M
      J. Neurochem., 2010;114(3):810-9.
      Species: Rat
      Sample Type: Cell Culture Supernates
    6. Mechanism of hepatoprotection in proestrus female rats following trauma-hemorrhage: heme oxygenase-1-derived normalization of hepatic inflammatory responses.
      Authors: Yang S, Hu S, Chen J, Choudhry MA, Rue LW, Bland KI, Chaudry IH
      J. Leukoc. Biol., 2009;85(6):1015-26.
      Species: Rat
      Sample Type: Serum
    7. Antithrombin inhibits bronchoalveolar activation of coagulation and limits lung injury during Streptococcus pneumoniae pneumonia in rats.
      Authors: Choi G, Hofstra JJ, Roelofs JJ, Rijneveld AW, Bresser P, van der Zee JS, Florquin S, van der Poll T, Levi M, Schultz MJ
      Crit. Care Med., 2008;36(1):204-10.
      Species: Rat
      Sample Type: BALF
    8. Natural anticoagulants limit lipopolysaccharide-induced pulmonary coagulation but not inflammation.
      Authors: Choi G, Vlaar AP, Schouten M, Van't Veer C, van der Poll T, Levi M, Schultz MJ
      Eur. Respir. J., 2007;30(3):423-8.
      Species: Rat
      Sample Type: Tissue Homogenates
    9. ACE mediates ventilator-induced lung injury in rats via angiotensin II but not bradykinin.
      Authors: Wosten-van Asperen RM, Lutter R, Haitsma JJ, Merkus MP, van Woensel JB, van der Loos CM, Florquin S, Lachmann B, Bos AP
      Eur. Respir. J., 2007;31(2):363-71.
      Species: Rat
      Sample Type: BALF
    10. Anti-rat soluble IL-6 receptor antibody down-regulates cardiac IL-6 and improves cardiac function following trauma-hemorrhage.
      Authors: Yang S, Hu S, Choudhry MA, Rue LW, Bland KI, Chaudry IH
      J. Mol. Cell. Cardiol., 2007;42(3):620-30.
      Species: Rat
      Sample Type: Tissue Homogenates
    11. Mechanism of the nongenomic effects of estrogen on intestinal myeloperoxidase activity following trauma-hemorrhage: up-regulation of the PI-3K/Akt pathway.
      Authors: Yu HP, Hsieh YC, Suzuki T, Choudhry MA, Schwacha MG, Bland KI, Chaudry IH
      J. Leukoc. Biol., 2007;82(3):774-80.
      Species: Rat
      Sample Type: Tissue Homogenates
    12. Mechanism of IL-6-mediated cardiac dysfunction following trauma-hemorrhage.
      Authors: Yang S, Hu S, Hsieh YC, Choudhry MA, Rue LW, Bland KI, Chaudry IH
      J. Mol. Cell. Cardiol., 2006;40(4):570-9.
      Species: Rat
      Sample Type: Tissue Homogenates
    13. Maintenance of lung myeloperoxidase activity in proestrus females after trauma-hemorrhage: upregulation of heme oxygenase-1.
      Authors: Yu HP, Yang S, Hsieh YC, Choudhry MA, Bland KI, Chaudry IH
      Am. J. Physiol. Lung Cell Mol. Physiol., 2006;291(3):L400-6.
      Species: Rat
      Sample Type: Tissue Homogenates
    Expand to show all 13 Citations
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