MCP-1/CCL2, granzyme B, KC (R&D Systems, MN), and IFNγ, IL-6 (BD Biosciences, NJ) supernatant levels were determined by ELISA. MCP-1/CCL2 and KC levels were normalized to the total protein concentration of the corresponding group. granzyme B and IFNγ levels were normalized to the number of ECs at the initiation of co-culture.
Mcp 1 ccl2
Manufactured by R&D Systems
Sourced in United States, Germany
MCP-1/CCL2 is a protein that belongs to the chemokine family. It functions as a chemoattractant for monocytes and basophils.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 6 (il 6), by R&D Systems (9 mentions)
Tnf α, by R&D Systems (8 mentions)
Il 1β, by R&D Systems (4 mentions)
Rantes ccl5, by R&D Systems (2 mentions)
Flexstation 3 microplate reader, by Molecular Devices (2 mentions)
Hts fluoroblok 96 multiwell insert system chemotaxis chambers, by Corning (2 mentions)
Infinity kit, by Thermo Fisher Scientific (2 mentions)
Testosterone, by ALPCO (2 mentions)
Wako kit, by Fujifilm (2 mentions)
Stanbio liquicolor kit, by EKF Diagnostics (2 mentions)
Tnf α, by Thermo Fisher Scientific (2 mentions)
Ccl3 mip 1α, by R&D Systems (2 mentions)
Mouse anti mcp 1, by Thermo Fisher Scientific (2 mentions)
Mouse npy, by Merck Group (2 mentions)
Horse anti mouse, by Cell Signaling Technology (2 mentions)
Ma5 17040, by Thermo Fisher Scientific (2 mentions)
Rabbit anti ucp1, by Cell Signaling Technology (2 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (2 mentions)
Mab1501, by Merck Group (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
30 protocols using mcp 1 ccl2
1
Quantifying Cytokine Levels in Co-cultures
2
Multiplex Cytokine Analysis of ADCC Supernatants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Supernatants of ADCC experiments (stored at −20 °C, see above) were used for protein analysis. A custom-made multiplex assay for CXCL13, Eotaxin/CCL11, G-CSF, GM-CSF, IFN-γ, IL-1β, IL-10, IL-12p70, IL-2, IL-23, IL-6, IL-8, IP10/CXCL10, MCP-1/CCL2, MIP1α/CCL3, MIP1β/CCL4, RANTES/CCL5 and TNF-α from R&D Systems (Minneapolis, MN, USA) was performed using LUMINEX technology (LX200™, Millipore, Burlington, MA) according to the manufacturer’s protocol. The cytokines IFN-γ, TGF-β and TNF-α (eBioscience, San Diego, CA, USA, cat. no. 88-7316-22, 88-8350-88, 39-8329, respectively) and granzyme B (MABTECH, Stockholm, Sweden, cat. no. 3485-3-250/6-250) were separately analyzed in sandwich ELISAs according to the manufacturer’s protocol.
3
Transendothelial Migration Assay with CB2 Agonist
4
Plasma Biomarkers in Fasting Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fasting (5 h) blood samples were taken from the tail vein to prepare EDTA plasma. Blood glucose was determined during blood sampling using a hand-held glucometer (Freestyle Freedom Light, Abbott Laboratories, Lake Bluff, IL, United States). Plasma insulin, leptin, adiponectin, cholesterol, triacylglycerols, ALT, CK-18M30 were determined as described (16 (link)). ELISA assays were used to measure plasma serum amyloid A (SAA; KMA0021 Thermo Fisher Scientific, Waltman, United States), MCP-1/CCL2 (DY479 R&D, Minneapolis, United States), MIF (DY1978 R&D, Fetuin A (DY1563 R&D) and LPS binding protein (LBP; HK205-02 Hycult Biotech, Uden, Netherlands) according to manufacturer’s protocol.
Lipoprotein profiles were analyzed by first separating lipoproteins into fractions using fast protein liquid chromatography (FPLC) with an AKTA apparatus (Pharmacia, Roosendaal, Netherlands), as previously described (17 (link), 18 (link)). Subsequently, total cholesterol and triacylglycerols were measured in the fractions collected for profiling with enzymatic assays (Roche diagnostics, Basel, CHF) (19 (link)–21 (link)).
Lipoprotein profiles were analyzed by first separating lipoproteins into fractions using fast protein liquid chromatography (FPLC) with an AKTA apparatus (Pharmacia, Roosendaal, Netherlands), as previously described (17 (link), 18 (link)). Subsequently, total cholesterol and triacylglycerols were measured in the fractions collected for profiling with enzymatic assays (Roche diagnostics, Basel, CHF) (19 (link)–21 (link)).
5
Chemotactic migration of THP-1 cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
THP-1 cells stably expressing either GPSM3-knockdown shRNA or scrambled shRNA control were serum-starved in HBSS + 1% BSA at 37°C for one hour and labeled with calcein-AM cell permeant fluorescent dye (494 nm/517 nm). Cells were washed and suspended in HBSS + 1% BSA + 10 mM HEPES, and 5 × 107 cells/ml were loaded into the upper well of a Falcon™ HTS FluoroBlok 96-Multiwell Insert System chemotaxis chambers (Corning, NY). Cells were allowed to migrate toward vehicle or 100 ng/mL MCP-1/CCL2 (R&D Systems, Minneapolis, MN) in the lower chamber. Fluorescence was measured at 2 minute intervals over the course of 60 minutes at 37°C using a Flexstation 3 microplate reader (Molecular Devices, Sunnyvale, CA). All data are representative of biological triplicates with technical replicates (i.e., n = 6).
6
Cytokine Profiling of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of IL-6, TNF-α (25–900-K21, Peprotech, Rocky Hill, NJ, USA), Growth-regulated oncogene (Gro)-α/CXCL1, IL-8/CXCL8, IL-10, monocyte chemoattractant protein 1 (MCP)1/CCL2, Macrophage-derived chemokine (MDC)/CCL22, pulmonary and activation-regulated chemokine (PARC)/CCL18, and thymus and activation regulated chemokine (TARC)/CCL17 (all from R&D Systems) in the culture supernatants were measured using commercial ELISA kits, according to the manufacturer's instructions. The supernatant was diluted to immediately prior to experiments as described in Table 2
Dilution used for specific cytokines for each conditions.
| Cytokine | MDM | Monocytes | ||||||
|---|---|---|---|---|---|---|---|---|
| Control | CSE 4% | LPS | LPS + CS4% | IL-4 | IL-4 + CS4% | Control | LPS | |
| Dilution | Dilution | |||||||
| IL-6 | 1:1 | 1:1 | 1:200 | 1:200 | 1:1 | 1:1 | 1:2 | 1:50 |
| TNF-α | 1:1 | 1:1 | 1:100 | 1:100 | 1:1 | 1:1 | 1:1 | 1:10 |
| IL-8/CXCCL8 | 1:10 or 1:100 | 1:100 | 1:100 or 1:200 | 1:100 | 1:100 | 1:100 | 1:100 | 1:100 |
| Gro-α/CXCL1 | 1:1 | 1:1 | 1:100 | 1:100 | 1:1 | 1:1 | 1:2 | 1:10 |
| MCP1/CCL2 | 1:4 | 1:4 | 1:20 | 1:20 | 1:4 | 1:4 | - | - |
| MDC/CCL22 | 1:100 | 1:100 | 1:100 or 1:200 | 1:100 or 1:200 | 1:100 or 1:200 | 1:100 or 1:200 | 1:100 | 1:100 |
| IL-10 | 1:1 | 1:1 | 1:1 | 1:1 | 1:1 | 1:1 | - | - |
| PARC/CCL18 | 1:10 | 1:10 | 01:10 | 01:10 | 01:10 | 1:10 | - | - |
| TARC/CCL17 | 1:1 | 1:1 | 1:1 | 1:1 | 1:1 | 1:1 | 1:2 | 1:2 |
The supernatant was diluted to immediately prior to experiments.
7
Monocyte Migration Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Migration of monocytes was performed using a 5 µm-pore size trans-well system (Corning, US). The migration inserts containing 1 × 106 monocytes in 100 µL were placed into a 24-well plate containing 600 µL of either media alone, recombinant human YKL-39 (100 ng/ml, Sino Biological Inc, China) or MCP-1/CCL2 (100 ng/ml, R&D, Germany). The monocytes in the inserts were incubated for 1 hour or 3 hours. After migration, the cells on the trans-well membranes were fixed with 4% PFA and stained with Mayer's hemalum solution (Merck, Germany). The cell numbers per field were counted of 10 random fields on each membrane (bottom side). The number of transmigrated cells in the lower chamber was counted by Casy Model TT cell counter (OLS, Germany).
8
Cytokine Profiling of C2C12 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokine concentrations in C2C12 conditioned media (cell culture supernatants) were analyzed for IL-6 (R&D Systems, Bio-Techne, RDY406-05), MCP-1 (CCL2) (R&D Systems, Bio-Techne, DY479-05), and TNF-α (R&D Systems, Bio-Techne, DY410-05) following the manufacturer’s recommendations.
9
Serum and Liver Lipid Profiling
10
Measurement of Serum MCP-1/CCL2 Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum was acquired from the blood obtained by antecubital vein puncture at 8:00 am before breakfast. Venipuncture of all participants was performed by a senior nurse. Following extraction, blood samples for serum isolation were kept for 20 minutes at room temperature. Then, the blood was centrifuged at 1500 g for 10 min. Next, serum samples were aliquoted and kept at -80 °C before measurement. Serum MCP-1/CCL2 concentrations (R&D Systems, MN, USA) were investigated using commercially available kits according to the suppliers' instruction manuals. The detection range was 31.2-2,000 pg/mL. The intra-assay and inter-assay coefficient of variation for MCP-1/CCL2 were 4.9% and 5.9%, respectively. Each sample examination was repeated three times, and the average result was calculated.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!