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Mouse CCL8/MCP-2 DuoSet ELISA

Catalog #: DY790
11 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY790
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Mouse CCL8 / MCP-2 ELISA Standard Curve
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Product Details
Procedure
Citations (11)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse CCL8/MCP-2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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 mouse CCL8/MCP-2. The suggested diluent is suitable for the analysis of most cell culture supernate 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)

Scientific Data

N/A Mouse CCL8 / MCP-2 ELISA Standard Curve View Larger

Mouse CCL8 / MCP-2 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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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.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL8/MCP-2

MCP-2 is a CC chemokine subfamily monocyte chemotactic protein. MCP-2 also chemoattracts activated NK cells, CD4+ and CD8+ T lymphocytes, and eosinophils and can induce histamine secretion from basophils. MCP-2 shares 62% amino acid sequence identity with MCP-1 and 58% with MCP-3.

Entrez Gene IDs:
6355 (Human); 20307 (Mouse)
Alternate Names:
C-C motif chemokine 8; CCL8; chemokine (C-C motif) ligand 8; HC14; MCP2; MCP-2; member 8 (monocyte chemotactic protein 2); monocyte chemoattractant protein 2; SCYA10; small-inducible cytokine A8

Assay Procedure

GENERAL ELISA PROTOCOL

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 Mouse CCL8/MCP-2 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.

11 Citations: Showing 1 - 10
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  1. Mesenchymal stem cells enhance CCL8 expression by podocytes in lupus-prone MRL.Faslpr mice
    Authors: Kim, HS;Lee, HK;Kim, K;Ahn, GB;Kim, MS;Lee, TY;Son, DJ;Kim, Y;Hong, JT;Han, SB;
    Scientific reports
    Species: Transgenic Mouse
    Sample Types: Cell Culture Supernates
  2. LHPP-mediated inorganic pyrophosphate hydrolysis-driven lysosomal acidification in astrocytes regulates adult neurogenesis
    Authors: Sha, L;Li, J;Shen, H;Wang, Q;Meng, P;Zhang, X;Deng, Y;Zhu, W;Xu, Q;
    Cell reports
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. Pathogenic roles and therapeutic potential of the CCL8-CCR8 axis in a murine model of IgG4-related sialadenitis
    Authors: F Honda, H Tsuboi, Y Ono, S Abe, H Takahashi, K Ito, K Yamada, M Kawano, Y Kondo, K Asano, M Tanaka, M Malissen, B Malissen, I Matsumoto, T Sumida
    Arthritis Research & Therapy, 2021-08-14;23(1):214.
    Species: Mouse
    Sample Types: Serum
  4. Phagocytosis of Leptospira by leukocytes from mice with different susceptibility to leptospirosis and possible role of chemokines
    Authors: PLD Silva, F Lauretti-F, M Caldas de, SS Lima, AE Covarrubia, M De Franco, E Carvalho, PL Ho, RMA da Costa, EAL Martins, JB Da Silva
    BMC Microbiol., 2019-01-07;19(1):4.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Induction of the MCP chemokine cluster cascade in the periphery by cancer cell-derived CCL3
    Authors: Elena Farmaki
    Cancer Lett, 2016-12-29;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. A CCL8 gradient drives breast cancer cell dissemination
    Oncogene, 2016-05-16;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  7. Knockdown of the antiapoptotic Bcl-2 family member A1/Bfl-1 protects mice from anaphylaxis.
    Authors: Ottina E, Lyberg K, Sochalska M, Villunger A, Nilsson G
    J Immunol, 2014-12-29;194(3):1316-22.
    Species: Mouse
    Sample Types: Serum
  8. Allergic airway inflammation decreases lung bacterial burden following acute Klebsiella pneumoniae infection in a neutrophil- and CCL8-dependent manner.
    Authors: Dulek D, Newcomb D, Goleniewska K, Cephus J, Zhou W, Reiss S, Toki S, Ye F, Zaynagetdinov R, Sherrill T, Blackwell T, Moore M, Boyd K, Kolls J, Peebles R
    Infect Immun, 2014-06-23;82(9):3723-39.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. beta-arrestin 2 inhibits proinflammatory chemokine production and attenuates contact allergic inflammation in the skin.
    Authors: Gaffal E, Jakobs M, Glodde N, Schroder R, Kostenis E, Tuting T
    J Invest Dermatol, 2014-02-27;134(8):2131-7.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Cannabinoid 1 receptors in keratinocytes modulate proinflammatory chemokine secretion and attenuate contact allergic inflammation.
    Authors: Gaffal E, Cron M, Glodde N, Bald T, Kuner R, Zimmer A, Lutz B, Tuting T
    J Immunol, 2013-04-12;190(10):4929-36.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Tumor microenvironment-derived proteins dominate the plasma proteome response during breast cancer induction and progression.
    Authors: Pitteri SJ, Kelly-Spratt KS, Gurley KE
    Cancer Res., 2011-06-08;71(15):5090-100.
    Species: Mouse
    Sample Types: Plasma

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