Levels of Hsp70 in the serum and hippocampus were determined by enzyme‐linked immunosorbent assay (ELISA). The Hsp70 ELISA Kit (ab133060, Abcam) was used for assessing Hsp70 levels in the hippocampus. The Hsp70 High Sensitivity ELISA Kit (ab133061, Abcam) was used for assessing Hsp70 levels in mouse serum. The following kits were used for the detection of inflammatory cytokines in the hippocampus: Mouse IL‐6 ELISA kit (ab100713, Abcam) for IL‐6, Mouse IL‐1 beta ELISA Kit (ab197742) for IL‐1ß, and Mouse monocyte chemoattractant protein‐1 (MCP1) ELISA Kit (ab100722) for MCP1. ELISA was performed according to the manufacturer's instruction.
Mcp 1 elisa kit
Manufactured by Abcam
The MCP-1 ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of monocyte chemoattractant protein-1 (MCP-1) levels in biological samples. The kit utilizes specific antibodies to capture and detect MCP-1 in the sample.
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Lab products found in correlation
Mouse il 1 beta elisa kit, by Abcam (1 mentions)
Mouse il 6 elisa kit, by Abcam (1 mentions)
Hsp70 elisa kit, by Abcam (1 mentions)
Hsp70 high sensitivity elisa kit, by Abcam (1 mentions)
Acetic acid, by Corning (1 mentions)
Db100b, by R&D Systems (1 mentions)
Mb100b, by R&D Systems (1 mentions)
Endo iwr 1, by Bio-Techne (1 mentions)
S31 201, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Il 1β elisa kit, by Abcam (1 mentions)
Ngal elisa kit, by R&D Systems (1 mentions)
Tnf α elisa kit, by R&D Systems (1 mentions)
5 protocols using mcp 1 elisa kit
1
Quantification of Hippocampus and Serum Hsp70 Levels
2
Quantifying MCP-1 and TGF-β in Kidney Cell Secretome
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MCP-1 and TGF-β in 24-hour–conditioned medium from RPTECs or human kidney cells (HK2, ATCC, CRL-2190) were quantified using an MCP-1 ELISA kit (Abcam, ab100721) or a TGF-β1 ELISA kit (R&D Systems, DB100B or MB100B) following the manufacturer’s instructions. Briefly, cells were plated in 6-well plates (3 × 105 to 4 × 105 cells/well) in complete medium. Twenty-four hours later, cells were incubated with serum-free medium containing 20 mM acetic acid or collagen I (50 μg/mL in 20 mM acetic acid, Corning). In some experiments, cells were incubated with the DDR1 inhibitor Cmp-1 (3 μM) synthesized as previously described (16 (link)), or the WNT/β-catenin inhibitor IWR1-endo (30 μM, Tocris), or the STAT3 inhibitor S31-201 (10 μM, MilliporeSigma) for 30 minutes prior to 24 hours of treatment with acetic acid or collagen I.
3
Serum Ang II and MCP-1 ELISA Analysis
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Serum Ang II concentrations were determined using a commercially available Ang II ELISA kit (Shanghai Enzyme-linked Biotechnology Co. LTD), while serum monocyte chemoattractant protein-1 (MCP-1) concentrations were measured with a MCP-1 ELISA kit (Abcam biotechnology Co. LTD) in accordance with the manufacturer's instructions.
4
Chemotaxis Assay for Monocyte Migration
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HK-2 cells were seeded in a 6-well plate and treated with the experimental medium in the absence or presence of 3mM eATP, 3.2mM PA or eATP plus PA. After a 24 h incubation, culture supernatants were harvested. Microporous membrane (pore size 8 μm) transwell inserts (Millipore, Watford, UK) were used for the chemotaxis assay.
THP-1 cells in 200μl serum-free RPMI were added to the upper chamber, with 500μl above culture supernatants in the lower chamber. THP-1 cells were allowed to migrate for 2 h incubation at 37 °C， 5 % CO 2 ， and then the inserts were fixed and stained with crystal violet (Sigma). The non-migratory cells were removed before the membrane was mounted and the number of migratory cells was observed under a microscope (Carl Zeiss). The monocyte chemoattractant protein-1 (MCP-1) in the supernatants was detected by MCP-1 ELISA kit (Abcam) according to the manufacturer's instructions.
THP-1 cells in 200μl serum-free RPMI were added to the upper chamber, with 500μl above culture supernatants in the lower chamber. THP-1 cells were allowed to migrate for 2 h incubation at 37 °C， 5 % CO 2 ， and then the inserts were fixed and stained with crystal violet (Sigma). The non-migratory cells were removed before the membrane was mounted and the number of migratory cells was observed under a microscope (Carl Zeiss). The monocyte chemoattractant protein-1 (MCP-1) in the supernatants was detected by MCP-1 ELISA kit (Abcam) according to the manufacturer's instructions.
5
Cytokine and Biomarker Profiling
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The cultured medium of NRK-52E cells and the serum samples of the experimental mice were collected and analyzed using the MCP-1 ELISA kit (Abcam, Cat. No. ab208979) and IL-1β ELISA kit (Abcam, Cat. No. ab197742). The serum samples were also analyzed using TNF-α ELISA kit (R&D Systems, Cat. No. MTA00B) and neutrophil gelatinase-associated lipocalin (NGAL) ELISA kit (R&D Systems, Cat. No. MLCN20) in accordance with the manufacturer’s instructions.
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