Mouse ccl2 elisa kit

Manufactured by R&D Systems

Sourced in United States

The Mouse CCL2 ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed to measure the levels of mouse CCL2 (also known as Monocyte Chemoattractant Protein-1, MCP-1) in cell culture supernatants, serum, and plasma samples. The kit utilizes a specific antibody coated on a microplate to capture CCL2 from the samples, and a detection antibody is then used to quantify the bound CCL2 using a colorimetric reaction.

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ELISAs (144 citations) Mouse ELISA kits (102 citations) CCL2 ELISA kit (6 citations)

3 protocols using mouse ccl2 elisa kit

Cytokine Profiling in Murine Retinal Tissue

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Retinal tissue collected from mice were lysed in 100 µL of lysis buffer with protease inhibitor cocktail as previously described¹⁰ (link) and homogenized by a Mini Bead Beater (BiosSpec Products, Bartlesville, OK). After centrifuging at 12,000×g for 10 minutes, the supernatant was used to measure the levels of tumor necrosis factor-α (TNF-α), IL-1β, and chemokine (C-C motif) ligand 2 (CCL2) in the retinal tissue. The conditional medium was also used to determine these cytokines’ secretions using mouse TNF-α ELISA kit (R&D, Minneapolis, MN), mouse IL-1β ELISA kit (R&D), mouse IL-6 ELISA kit (R&D), and mouse CCL2 ELISA kit (R&D), respectively, following the manufacturer's instructions. Serum creatine and blood urea nitrogen were measured using SCR ELISA kit (cat# YS04392B, YaJi Biological, China) and blood urea nitrogen ELISA kit (cat# C013-2, YaJi Biological, China) respectively.

Liu X., Xu B, & Gao S. (2021). Spleen Tyrosine Kinase Mediates Microglial Activation in Mice With Diabetic Retinopathy. Translational Vision Science & Technology, 10(4), 20.

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ELISA Quantification of Mouse CCL2

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Mouse CCL2 ELISA kit was purchased from R&D systems. For in vitro experiments, cultured cells were collected after treatment. For in vivo experiments, animals were transcardially perfused with PBS. The L5 spinal cord was dissected. Astrocytes and spinal cord tissues were homogenized in a lysis buffer containing protease and phosphatase inhibitors (Sigma, St Louis, MO). Protein concentrations were determined by BCA Protein Assay (Pierce, Rockford, IL). For each reaction in a 96-well plate, 100 µg of proteins were used, and ELISA was performed according to manufacturer’s protocol. The standard curve was included in each experiment.

Lu Y., Jiang B.C., Cao D.L., Zhang Z.J., Zhang X., Ji R.R, & Gao Y.J. (2014). TRAF6 upregulation in spinal astrocytes maintains neuropathic pain by integrating TNF-α and IL-1β signaling. Pain, 155(12), 2618-2629.

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Microglia and Astrocyte Activation Assay

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Dulbecco's modified Eagle's medium (DMEM), 0.25% Trypsin-EDTA solution, fetal calf serum (FCS), penicillin/streptomycin and poly-D-lysine were purchased from Gibco-BRL (Grand Island, NY, USA). RS102895 was purchased from Sigma-Aldrich (St. Louis, MO, USA). RIPA buffer and the BCA kit were purchased from Beyotime (Shanghai, China). Fluoroshieldmounting medium with 4, 6-diamidino-2-phenylindole (DAPI) and mouse anti-OX42 monoclonal antibody were purchased from Abcam (Hongkong, China). Mouse CCL-2 ELISA kit and TNF-α were obtained from R&D Systems, Inc. (Minneapolis, MN, USA). Rabbit anti-Iba1 monoclonal antibodies and anti-GFAP polyclonal antibodies were purchased from Abcam (Hongkong, China). A FITC-conjugated goat anti-rabbit IgG antibody was purchased from Santa Cruz (Santa Cruz Biotechnology, USA). Rat anti-mouse CD86 was purchased from BD Bioscience (San Jose, CA, USA). Aleva Fluor 488-conjugated donkey anti-rat IgG was purchased from Invitrogen (Carlsbad, CA, USA).

He M., Dong H., Huang Y., Lu S., Zhang S., Qian Y, & Jin W. (2016). Astrocyte-Derived CCL2 is Associated with M1 Activation and Recruitment of Cultured Microglial Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 38(3).

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