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R&D Systems DuoSet ELISAs
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Rat JE/MCP-1/CCL2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Range
7.8 - 500 pg/mL
Sufficient Materials
For five 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat Monocyte Chemotactic Protein 1 (MCP-1). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Rat JE/MCP-1/CCL2 DuoSet ELISA

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.

14 Citations: Showing 1 - 10
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  1. Pb Induces MCP-1 in the Choroid Plexus
    Authors: H Gu, Y Xu, N Du, Y Yu, W Zheng, Y Du
    Biology, 2022;11(2):.
    Species: Rat
    Sample Types: Cell Culture Supernates
  2. Effects of a lysophosphatidic acid receptor 1 antagonist on hypertensive renal injury in Dahl-Iwai salt-sensitive rats
    Authors: T Naruse, H Otake, T Takahashi
    Oncogene, 2022;149(4):179-188.
    Species: Rat
    Sample Types: Serum
  3. Mechanosensitive cation channel Piezo1 contributes to ventilator-induced lung injury by activating RhoA/ROCK1 in rats
    Authors: Y Zhang, L Jiang, T Huang, D Lu, Y Song, L Wang, J Gao
    Respiratory Research, 2021;22(1):250.
    Species: Rat
    Sample Types: BALF
  4. Hydrogen Sulfide Antagonizes Sleep Deprivation-induced Depression- and Anxiety-like Behaviors by Inhibiting Neuroinflammation in A Hippocampal Sirt1-dependent Manner
    Authors: X Kang, L Jiang, F Lan, YY Tang, P Zhang, W Zou, YJ Chen, XQ Tang
    Brain research bulletin, 2021;177(0):194-202.
    Species: Rat
    Sample Types: Tissue Homogenates
  5. Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1
    Authors: E D'Alessand, B Scaf, C Munts, A van Hunnik, CJ Trevelyan, S Verheule, HMH Spronk, NA Turner, H Ten Cate, U Schotten, FA van Nieuwe
    Cells, 2021;10(11):.
    Species: Rat
    Sample Types: Cell Culture Supernates
  6. Study of the Role of the Tyrosine Kinase Receptor MerTK in the Development of Kidney Ischemia-Reperfusion Injury in RCS Rats
    Authors: T Pelé, S Giraud, S Joffrion, V Ameteau, A Delwail, JM Goujon, L Macchi, T Hauet, F Dkhissi, O Benzakour
    International Journal of Molecular Sciences, 2021;22(22):.
    Species: Rat
    Sample Types: Plasma
  7. Inhibition of proprotein convertase subtilisin/kexin type 9 attenuates 2,4,6-trinitrobenzenesulfonic acid-induced colitis via repressing toll-like receptor 4/nuclear factor-kappa B
    Authors: L Lei, X Li, YJ Yuan, ZL Chen, JH He, JH Wu, XS Cai
    Kaohsiung J. Med. Sci., 2020;0(0):.
    Species: Rat
    Sample Types: Tissue Culture Supernates
  8. Inhibition of HtrA2 alleviates inflammatory response and cell apoptosis in lipopolysaccharide?induced acute pneumonia in rats
    Authors: X Wang
    Molecular Medicine Reports, 2020;0(0):.
    Species: Rat
    Sample Types: Cell Culture Supernates
  9. NMR-based metabolomics approach reveals effects of antioxidant nutrients in sepsis-induced changes in rat liver injury
    Authors: D Xu, S Liao, Y Lv, J Wang, L Kong
    J. Nutr. Biochem., 2020;85(0):108440.
    Species: Rat
    Sample Types: Serum
  10. Resveratrol Effects on a Diabetic Rat Model with Coronary Heart Disease
    Authors: X Huo, T Zhang, Q Meng, C Li, B You
    Med. Sci. Monit., 2019;25(0):540-546.
    Species: Rat
    Sample Types: Serum
  11. Nrf2 Suppression Delays Diabetic Wound Healing Through Sustained Oxidative Stress and Inflammation
    Authors: M Li, H Yu, H Pan, X Zhou, Q Ruan, D Kong, Z Chu, H Li, J Huang, X Huang, A Chau, W Xie, Y Ding, P Yao
    Front Pharmacol, 2019;10(0):1099.
    Species: Rat
    Sample Types: Serum
  12. A Rat Model of Maternal Polycystic Ovary Syndrome Shows that Exposure to Androgens In Utero Results in Dysbiosis of the Intestinal Microbiota and Metabolic Disorders of the Newborn Rat
    Authors: T Gulan, T Yeernuer, S Sui, N Mayinuer
    Med. Sci. Monit., 2019;25(0):9377-9391.
    Species: Rat
    Sample Types: Serum
  13. Overexpression of IL-10 Enhances the Efficacy of Human Umbilical-Cord-Derived Mesenchymal Stromal Cells in E. coli Pneumosepsis
    Authors: M Jerkic, C Masterson, L Ormesher, S Gagnon, S Goyal, R Rabani, G Otulakowsk, H Zhang, BP Kavanagh, JG Laffey
    J Clin Med, 2019;8(6):.
    Species: Rat
    Sample Types: Bronchoalveolar lavage fluid
  14. Maternal Consumption of Low-Isoflavone Soy Protein Isolate Confers the Increased Predisposition to Alcoholic Liver Injury in Adult Rat Offspring
    Authors: SB Won, YH Kwon
    Nutrients, 2018;10(3):.
    Species: Rat
    Sample Types: Serum


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