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hCCL2/MCP-1 QKit (1 Kit)
品牌： R&D Systems
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Sample Values
Serum/Plasma/Urine - Samples from apparently healthy volunteers were evaluated for the presence of human MCP-1 in this assay. No medical histories were available for the donors used in this study. The reported urine values are actual and are not normalized for creatinine content.| Sample Type | Mean (pg/mL) | Range (pg/mL) |
| Serum (n=37) | 370 | 200-722 |
| EDTA plasma (n=37) | 153 | 72-295 |
| Citrate plasma (n=37) | 196 | 134-436 |
| Heparin plasma (n=37) | 242 | 113-340 |
| Urine (n=37) | 211 | 42-410 |
Cell Culture Supernates - Human peripheral blood leukocytes were cultured in RPMI 1640 and supplemented with 10% fetal bovine serum. The cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 2 or 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of human MCP-1.
| Condition | Day 2 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 647 | 1785 |
| Stimulated | 67,225 | 70,000 |
Recovery
The recovery of MCP-1 spiked to three levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 96 | 88-107 |
| Citrate Plasma (n=5) | 100 | 94-107 |
| EDTA Plasma (n=5) | 96 | 92-102 |
| Heparin Plasma (n=5) | 102 | 94-114 |
| Serum (n=5) | 103 | 92-113 |
| Urine (n=5) | 92 | 85-100 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of MCP-1 and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.
- Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process twice for a total of 3 washes.
- Add 200 µL of Conjugate to each well.
- For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
For Cell Culture Supernate & Urine Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour. - Aspirate and wash 3 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent for Serum & Plasma Samples Only
200 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Human CCL2/MCP-1 Quantikine ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (200 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL), Urine (100 uL)
Sensitivity
10 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
Specificity
Natural and recombinant human MCP-1
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
背景
别名
C-C motif chemokine ligand 2,CCL2,GDCF-2,HC11,HSMCR30,MCAF,Mcp1,MCP-1,SCYA2,SMC-CF
背景
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)
Alternate Names:
C-C motif chemokine ligand 2; CCL2; GDCF-2; HC11; HSMCR30; MCAF; Mcp1; MCP-1; SCYA2; SMC-CF
研究领域
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