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Product Info Summary
| SKU: | EK0441 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine. |
Product info
Product Name
Human MCP-1 / CCL2 ELISA Kit PicoKine®
SKU/Catalog Number
EK0441
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human MCP-1 / CCL2 ELISA Kit PicoKine® (96 Tests). Quantitate Human CCL2 in cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine. Sensitivity: 1pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Human MCP-1 / CCL2 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0441)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: Q24-T99
Sensitivity
<1 pg/ml
Assay Range
15.6 pg/ml - 1,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0441 for more details
Cross-reactivity
There is cross-reactivity with human Eotaxin, MCP-3<1%.
Reactive Species
EK0441 is reactive to CCL2 in Human samples
Validated Sample Types
cell culture supernatants, serum, plasma (heparin, EDTA, citrate) and urine.
Application Guarantee
EK0441 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of CCL2
Monocyte chemoattractant protein-1 (MCP-1), a member of the chemokine (chemotactic cytokine) family, is a potent monocyte agonist that is upregulated by oxidized lipids. MCP-1 is also known as CCL2, SCYA2, MCAF. MCAF is a member of family of factors involved in immune and inflammatory responses. The amino acid sequence deduced from the nucleotide sequence reveals the primary structure of the MCAF precursor to be composed of a putative signal peptide sequence of 23 amino acid residues and a mature MCAF sequence of 76 amino acid residues. MCP-1 plays a unique and crucial role in the initiation of atherosclerosis and may provide a new therapeutic target in this disorder. Human MCP-1 is a 8.7KDa non-glycoprotein, consisting of 99 amino acids in precursor form and 76 amino acids in mature form.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0441-CAP | Anti-Human CCL2 Pre-coated 96-well strip microplate | 1 |
| EK0441-ST | Human CCL2 Standard | 2 vials, 10 ng/tube |
| EK0441-DA | Human CCL2 Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.015 | 0.075 | 0.126 | 0.25 | 0.459 | 0.875 | 1.463 | 2.275 |
Data Example Images
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Human MCP-1 PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect CCL2, Dilution ratio of 1:5, concentration in serum and plasma is around 500 pg/ml..
Some articles we found to cite concentrations of CCL2 in samples: 23629827 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human MCP-1 / CCL2 ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| 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% |
Specific Publications For Human MCP-1 / CCL2 ELISA Kit PicoKine® (EK0441)
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Customer Reviews
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Customer Q&As
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8 Customer Q&As for Human MCP-1 / CCL2 ELISA Kit PicoKine®
Question
Q: which procedure should I follow in order to thaw whole blood sample for CCL2 ELISA after freezing?
C. Mitchell
Verified customer
Asked: 2020-09-20
Answer
A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test CCL2 for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.
Boster Scientific Support
Answered: 2020-09-20
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2020-03-11
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are included in the product insert. Components in lysate and lysis buffer may affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2020-03-11
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
L. Lopez
Verified customer
Asked: 2019-04-02
Answer
A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2019-04-02
Question
Q: What is the optimal O.D. value for CCL2 ELISA kit? I used your CCL2 ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain CCL2 even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2018-05-15
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. regarding your protocol, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. normally, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by changeing cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this CCL2 ELISA kit to have sensitivity of 1pg/ml, that means the minimum amount of CCL2 that can be declared/interpreted as positive by the above standard is 1pg/ml.
Boster Scientific Support
Answered: 2018-05-15
Question
Q: how to proceed with the analysis of ELISA data? I measured CCL2 level in plasma.
Verified Customer
Verified customer
Asked: 2018-04-19
Answer
A: we recommend you this article on ELISA data analysis. bosterbio.com/elisa-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/elisa-data-analysis-online
Boster Scientific Support
Answered: 2018-04-19
Question
Q: can I use citrate plasma as samples in Human CCL2 Picokine® ELISA Kit (Catalog # EK0441)?
Verified Customer
Verified customer
Asked: 2018-04-02
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can sequester metal ions from the functional domain of CCL2 causing disruption of its protein structure. CCL2 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the CCL2 ELISA, treating samples with a number of anticoagulants and decided that heparin, EDTA or citrate can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2018-04-02
Question
Q: if the enzyme conjugated CCL2 antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the CCL2 antigen?
Verified Customer
Verified customer
Asked: 2017-01-30
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2017-01-30
Question
What serum dilution should be used for this antibody EK0441? I will be using human serum samples.
Verified Customer
Verified customer
Asked: 2013-09-23
Answer
We recommend to assay the human serum samples neat with standard buffer. The serum must be 2 fold dilution.
Boster Scientific Support
Answered: 2013-09-23
