| Field | Specification |
|---|---|
| Target | |
| Alternative names | C-C motif chemokine 2;HC11;Monocyte chemoattractant protein 1;Monocyte chemotactic and activating factor;MCAF;Monocyte chemotactic protein 1;MCP-1;Monocyte secretory protein JE;Small-inducible cytokine A2;CCL2;MCP1, SCYA2 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: Q24-T99 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Urine |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This EZ-Set™ kit supplies a matched capture and detection antibody pair and a standard for building your own sandwich ELISA for human C-C motif chemokine 2 (gene CCL2) in cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine. You coat the plate with the capture antibody, add samples and standards, detect with the biotinylated antibody and an avidin–biotin–peroxidase complex, develop with TMB and read at 450 nm. Plates, diluents, wash buffer, TMB and stop solution are not included.
CCL2 (also called MCP-1) is a C-C chemokine that recruits monocytes, memory T cells and basophils by binding the receptor CCR2. It is induced at sites of inflammation and tissue injury and is one of the most studied chemokines in atherosclerosis, obesity, fibrosis, neuroinflammation and the tumor microenvironment.
Kit Components
| Rabbit anti-human CCL2 polyclonal antibody (Capture Antibody) | 500 µL, 0.4 mg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-human CCL2 polyclonal antibody (Detection Antibody) | 500 µL, 5 µg/mL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant human CCL2 standard | 3 × 10 ng/tube |
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Standard | Recombinant human CCL2 (Gln24–Thr99), expressed in E. coli |
| Cross-reactivity | Below 1% cross-reactivity with human Eotaxin, MCP-3 |
| Intra-assay CV | 5.0–6.9% (3 samples, n = 16 each) |
| Inter-assay CV | 6.6–7.8% (3 samples, n = 24 each) |
| Format | Reagents for five 96-well plates |
Example standard curve (OD450, pg/mL): 0 = 0.008; 15.6 = 0.1; 31.2 = 0.206; 62.5 = 0.426; 125 = 0.826; 250 = 1.271; 500 = 1.762; 1000 = 2.087. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.008 | 0.1 | 0.206 | 0.426 | 0.826 | 1.271 | 1.762 | 2.087 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 31 | 159 | 477 | 30 | 148 | 481 |
| Standard deviation | 1.55 | 8.74 | 32.91 | 1.98 | 9.76 | 37.51 |
| CV (%) | 5% | 5.5% | 6.9% | 6.6% | 6.6% | 7.8% |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0441-CA | Rabbit anti- human CCL2 polyclonal antibody (Capture Antibody) | 500 µl, 0.4 mg/mL (recommended dilution 1:100) | Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. |
| EZ0441-DA | Biotinylated goat anti- human CCL2 polyclonal antibody (Detection Antibody) | 500 µl, 5 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0441-ST | Lyophilized recombinant human CCL2 standard | 10 ng/tube×3 | Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. |
Materials required but not provided
- Microplate reader in standard size.
- Automated plate washer.
- Incubator.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- 96 well microplate (Cat# AR1100)
- Plate Sealers.
- Capture Antibody Diluent: PBS.
- Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered.
- Substrate Solution: Tetramethylbenzidine (Cat# AR1104)
- Stop Solution: 2 N H2SO4 (Cat# AR1105)
- Wash Buffer (PBS and PBS-T). Recipes:
- PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 µm filtered.
- PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4.
- *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
Can’t Find What You’re Looking For? We can help you source the best match or customize an ELISA solution for your study. Options may include alternative target synonyms, different species reactivity, sample type/matrix compatibility (serum/plasma/lysate/supernatant), assay format (sandwich/competitive), sensitivity/range, detection chemistry (colorimetric/fluorescent/chemiluminescent), plate format (pre-coated/uncoated, strips vs full plate), and bulk or custom packaging. Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
Alicja Bauer et al. (2024) Circulating Monocyte Chemoattractant Protein-1 (MCP-1) in Patients with Primary Biliary Cholangitis. International Journal of Molecular Sciences. 10.3390/ijms25021333
Fangfang Ding et al. (2024) Polystyrene microplastics with absorbed nonylphenol induce intestinal dysfunction in human Caco-2 cells. Environmental Toxicology and Pharmacology. 10.1016/j.etap.2024.104426
Joana Viegas et al. (2024) A Novel Bionebulizer Approach to Study the Effects of Natural Mineral Water on a 3D In Vitro Nasal Model from Allergic Rhinitis Patients. Biomedicines. 10.3390/biomedicines12020408
