Rat CXCL3/CINC-2 alpha/beta Quantikine ELISA Kit

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RCN200
Rat CXCL3/GRO gamma/CINC-2/DCIP-1 ELISA Standard Curve
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Rat CXCL3/CINC-2 alpha/beta 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
3.9 pg/mL
Assay Range
6.3 - 400 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant rat CINC-2 alpha/beta
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-2 alpha/beta Immunoassay is a 4.5 hour solid-phase ELISA designed to measure rat CINC-2 alpha/beta in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat CINC-2 alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant rat CINC-2 alpha and recombinant rat CINC-2 beta. Results obtained using natural rat CINC-2 aalph/beta 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-2 alpha/beta.

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
Sample 1 2 3 1 2 3
n 20 20 20 47 47 47
Mean (pg/mL) 10.2 52.4 217 13 48.1 215
Standard Deviation 1.1 2.6 7.3 1.9 3.8 8.3
CV% 10.8 5 3.4 14.6 7.9 3.9

Recovery

The recovery of rat CINC-2 alpha/beta spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 101 98-107
EDTA Plasma (n=6) 99 90-109
Heparin Plasma (n=6) 112 103-119
Serum (n=6) 97 93-102

Linearity

To assess the linearity of the assay, samples containing rat CINC-2 alpha/beta in each matrix were diluted with Calibrator Diluent and then assayed.
Rat CXCL3/GRO gamma/CINC-2/DCIP-1 ELISA Linearity

Scientific Data

Rat CXCL3/GRO gamma/CINC-2/DCIP-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: CXCL3/GRO gamma/CINC-2/DCIP-1

CXCL3/GRO gamma, also known as DCIP-1 in mouse and CINC-2 in rat, is a member of the CXC chemokine family. Human CXCL3/GRO gamma is 107 amino acids (aa) in length, which includes a proposed signal peptide (aa 1–34), with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 65% and 59% aa sequence identity with the human protein, respectively. Genome-wide analysis suggests that the protein encoded by the mouse Cxcl1 gene is homologous to human CXCL3/GRO gamma protein. CXCL3/GRO gamma is chemotactic for granulocytes, including neturophils.

Entrez Gene IDs:
2921 (Human); 330122 (Mouse); 171551 (Rat)
Alternate Names:
chemokine (C-X-C motif) ligand 3; CINC-2; CINC-2b; C-X-C motif chemokine 3; CXCL3; Dcip1; Gm1960; GRO gamma; GRO3 oncogene; GRO3; GROG; GRO-gamma; GRO-gamma(1-73); Growth-regulated protein gamma; Macrophage inflammatory protein 2-beta; melanoma growth stimulatory activity gamma; MGSA gamma; MIP2B; MIP-2b; MIP2-beta; SCYB3
&#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. 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 for Rat CXCL3/CINC-2 alpha/beta 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.

4 Citations: Showing 1 - 4
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  1. Effect of Different Molecular Weights of Polyacrylic Acid on Rat Lung Following Intratracheal Instillation
    Authors: C Nishida, H Izumi, T Tomonaga, KY Wang, H Higashi, JI Takeshita, R Ono, K Sumiya, S Fujii, Y Hata, K Sakurai, Y Higashi, K Yamasaki, K Yatera, Y Morimoto
    International Journal of Molecular Sciences, 2022-09-07;23(18):.
    Species: Rat
    Sample Types: BALF
  2. Impaired integrin &alpha5 /&beta1 -mediated hepatocyte growth factor release by stellate cells of the aged liver
    Authors: F Rohn, C Kordes, T Buschmann, D Reichert, M Wammers, G Poschmann, K Stühler, AS Benk, F Geiger, JP Spatz, D Häussinger
    Aging Cell, 2020-03-11;0(0):e13131.
    Species: Rat
    Sample Types: Cell Culture Supernates
  3. 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, 2017-01-16;2017(0):4245309.
    Species: Mouse
    Sample Types: Tissue Homogenates
  4. Comprehensive gene expression profiling of rat lung reveals distinct acute and chronic responses to cigarette smoke inhalation.
    Authors: Stevenson CS, Docx C, Webster R, Battram C, Hynx D, Giddings J, Cooper PR, Chakravarty P, Rahman I, Marwick JA, Kirkham PA, Charman C, Richardson DL, Nirmala NR, Whittaker P, Butler K
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007-08-24;293(5):L1183-93.
    Species: Rat
    Sample Types: BALF

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