• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Sample Type & Volume Required

  100 µL

• Range

  3.91 - 250 pg/mL

• Sufficient Materials

  For five or fifteen 96-well plates*

• Specificity

  Please see the

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

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse CCL2/JE. 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 # ) 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 # ), 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 # ), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # ), 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 # )

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

Preparation and Storage

• 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);

• Aliases:

  CCL2; MCAF; MCP-1