CCL3/MIP-1 alpha, is a beta chemokine that can be induced in a variety of hematopoietic cells, fibroblasts, smooth muscle cells, and epithelial cells. It forms complexes with sulfated proteoglycans and also associates into noncovalently-linked dimers and high molecular weight polymers. These complexes of CCL3 are protected from proteolytic digestion by insulin degrading enzyme (IDE) which can cleave the monomeric chemokine. CCL3 signals through CCR1, CCR3, and CCR5, and it is internalized by the decoy chemokine receptor D6. CCL3 promotes the chemoattraction, adhesion to activated vascular endothelium, and cellular activation of many hematopoietic cell types. It can promote tumor metastasis, hyperalgesia, and inflammation and suppress HIV replication and hematopoietic stem cell proliferation.
Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
R&D Systems | Catalog # MMA00
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL)
Sensitivity
1.5 pg/mL
Assay Range
4.7-300 pg/mL (Cell Culture Supernates, Serum)
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Product Summary for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
The Quantikine Mouse MIP-1 alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MIP-1 alpha levels in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse MIP-1 alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring mouse MIP-1 alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of natural mouse MIP-1 alpha.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Mouse
Specificity
Natural and recombinant mouse MIP-1 alpha
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.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 13.7 | 86 | 184 | 13.8 | 83 | 179 |
| Standard Deviation | 0.6 | 3.1 | 5.0 | 0.9 | 5.2 | 12 |
| CV% | 4.4 | 3.6 | 2.7 | 6.5 | 6.3 | 6.7 |
Recovery for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
The recovery of mouse MIP-1 alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Supernates (n=5) | 105 | 95-114 |
| Serum (n=9) | 97 | 85-108 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse MIP-1 alpha in each matrix were diluted with Calibrator Diluent and then assayed.
Scientific Data Images for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
Mouse CCL3/MIP-1 alpha ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CCL3/MIP-1 alpha
Alternate Names
G0S19-1, LD78a, MIP-1 alpha, MIP1 alpha, MIP1A, PAT 464.1, SCYA3, SIS-beta
Entrez Gene IDs
Gene Symbol
CCL3
Additional CCL3/MIP-1 alpha Products
Product Documents for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Citations for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit
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Sample Tested: Cell culture supernatantVerified Customer | Posted 02/07/2020
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Sample Tested: Purified Standard and 3T3-L1 mouse embryonic fibroblast adipose-like cell lineVerified Customer | Posted 01/05/2018
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Sample Tested: Cell Culture SupernatesVerified Customer | Posted 10/14/2016
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Protocols
View specific protocols for Mouse CCL3/MIP-1 alpha Quantikine ELISA Kit (MMA00):
Refer to the product for complete assay procedure.
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.
- Add 50 µL of Assay Diluent to each well.
- Add 50 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 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 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
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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