Human Cell Biology ELISA Kits 3
Human MCP-3 (Monocyte Chemotactic Protein 3) CLIA Kit (HUES00024)
- SKU:
- HUES00024
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 4.69pg/mL
- Range:
- 7.81-500pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- MCP3, FIC, MARC, NC28, SCYA6, SCYA7
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 7.81-500 pg/mL |
| Sensitivity: | 4.69 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human MCP-3 in samples. No significant cross-reactivity or interference between Human MCP-3 and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human MCP-3. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MCP-3 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MCP-3, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human MCP-3. The concentration of Human MCP-3 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | CCL7: Chemotactic factor that attracts monocytes and eosinophils, but not neutrophils. Augments monocyte anti-tumor activity. Also induces the release of gelatinase B. This protein can bind heparin. Binds to CCR1, CCR2 and CCR3. Belongs to the intercrine beta (chemokine CC) family. |
| UniProt Protein Details: | Protein type:Secreted; Motility/polarity/chemotaxis; Secreted, signal peptide Chromosomal Location of Human Ortholog: 17q11. 2-q12 Cellular Component: extracellular space; extracellular region Molecular Function:heparin binding; CCR1 chemokine receptor binding; CCR2 chemokine receptor binding; chemokine activity Biological Process: cellular calcium ion homeostasis; regulation of cell shape; monocyte chemotaxis; cell-cell signaling; eosinophil chemotaxis; response to gamma radiation; cytoskeleton organization and biogenesis; immune response; chemotaxis; inflammatory response; signal transduction; positive regulation of cell migration |
| NCBI Summary: | This gene encodes monocyte chemotactic protein 3, a secreted chemokine which attracts macrophages during inflammation and metastasis. It is a member of the C-C subfamily of chemokines which are characterized by having two adjacent cysteine residues. The protein is an in vivo substrate of matrix metalloproteinase 2, an enzyme which degrades components of the extracellular matrix. This gene is part of a cluster of C-C chemokine family members on chromosome 17q. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P80098 |
| NCBI GenInfo Identifier: | 1170890 |
| NCBI Gene ID: | 6354 |
| NCBI Accession: | P80098. 3 |
| UniProt Secondary Accession: | P80098,Q569J6, |
| UniProt Related Accession: | P80098 |
| Molecular Weight: | 11,200 Da |
| NCBI Full Name: | C-C motif chemokine 7 |
| NCBI Synonym Full Names: | chemokine (C-C motif) ligand 7 |
| NCBI Official Symbol: | CCL7 |
| NCBI Official Synonym Symbols: | FIC; MARC; MCP3; NC28; MCP-3; SCYA6; SCYA7 |
| NCBI Protein Information: | C-C motif chemokine 7; small-inducible cytokine A7; monocyte chemotactic protein 3; monocyte chemoattractant protein 3; small inducible cytokine A7 (monocyte chemotactic protein 3) |
| UniProt Protein Name: | C-C motif chemokine 7 |
| UniProt Synonym Protein Names: | Monocyte chemoattractant protein 3; Monocyte chemotactic protein 3; MCP-3; NC28; Small-inducible cytokine A7 |
| Protein Family: | C-C motif chemokine |
| UniProt Gene Name: | CCL7 |
| UniProt Entry Name: | CCL7_HUMAN |
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | RLU | Average | Corrected |
| 500 | 51161 51913 | 51537 | 51508 |
| 250 | 18907 23089 | 20998 | 20969 |
| 125 | 9971 8821 | 9396 | 9367 |
| 62.5 | 4227 4797 | 4512 | 4483 |
| 31.25 | 2317 2281 | 2299 | 2270 |
| 15.63 | 1262 1238 | 1250 | 1221 |
| 7.81 | 725 755 | 740 | 711 |
| 0 | 29 29 | 29 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MCP-3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MCP-3 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 25.30 | 53.26 | 234.51 | 26.68 | 54.06 | 254.40 |
| Standard deviation | 3.14 | 5.48 | 16.51 | 2.24 | 4.00 | 17.43 |
| C V (%) | 12.41 | 10.29 | 7.04 | 8.40 | 7.40 | 6.85 |
Recovery
The recovery of Human MCP-3 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 87-99 | 92 |
| EDTA plasma (n=5) | 85-97 | 91 |
| Cell culture media (n=5) | 93-106 | 100 |
Linearity
Samples were spiked with high concentrations of Human MCP-3 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 102-117 | 98-111 | 88-99 |
| Average (%) | 108 | 105 | 93 | |
| 1:4 | Range (%) | 97-112 | 94-107 | 90-102 |
| Average (%) | 103 | 100 | 95 | |
| 1:8 | Range (%) | 98-114 | 89-104 | 101-116 |
| Average (%) | 106 | 95 | 107 | |
| 1:16 | Range (%) | 88-101 | 89-99 | 86-97 |
| Average (%) | 94 | 94 | 92 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro CLIA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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