Human CCL3/MIP-1 alpha Quantikine ELISA Kit

R&D Systems | Catalog # DMA00

R&D Systems
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Key Product Details

Assay Length

3.5 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (10 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Citrate Plasma (200 µL)

Sensitivity

10 pg/mL

Assay Range

31.2-1000 pg/mL (Cell Culture Supernates)
46.9-1500 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human CCL3/MIP-1 alpha Quantikine ELISA Kit

The Quantikine Human MIP-1 alpha Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure MIP-1 alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-1 alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring human 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 human 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

Human

Specificity

Natural and recombinant human 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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 116 442 726 53.6 313 623
Standard Deviation 3.3 5.9 14.0 3.7 13.4 25.7
CV% 2.8 1.3 1.9 6.9 4.3 4.1

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 140 688 1084 50.7 306 613
Standard Deviation 12.5 60.4 55.4 5.9 15.1 37.8
CV% 8.9 8.8 5.1 11.6 4.9 6.2

Recovery for Human CCL3/MIP-1 alpha Quantikine ELISA Kit

The recovery of MIP-1 alpha spiked to different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 100 88-117
Citrate Plasma (n=5) 94 85-104
EDTA Plasma (n=5) 96 81-109
Heparin Plasma (n=5) 98 82-121
Serum (n=5) 95 87-113

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of MIP-1 alpha in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human CCL3/MIP-1 alpha ELISA Linearity

Scientific Data Images for Human CCL3/MIP-1 alpha Quantikine ELISA Kit

Human CCL3/MIP-1 alpha ELISA Cell Culture Supernate Standard Curve

Human CCL3/MIP-1 alpha ELISA Cell Culture Supernate Standard Curve

Human CCL3/MIP-1 alpha ELISA Serum/Plasma Standard Curve

Human CCL3/MIP-1 alpha ELISA Serum/Plasma Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. 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

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.

Alternate Names

G0S19-1, LD78a, MIP-1 alpha, MIP1 alpha, MIP1A, PAT 464.1, SCYA3, SIS-beta

Entrez Gene IDs

6348 (Human); 20302 (Mouse); 25542 (Rat); 448787 (Canine); 102136134 (Cynomolgus Monkey)

Gene Symbol

CCL3

Additional CCL3/MIP-1 alpha Products

Product Documents for Human 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 Human 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 Human CCL3/MIP-1 alpha Quantikine ELISA Kit

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    Name: Anonymous
    Sample Tested: BMEC
    Verified Customer | Posted 07/11/2019
  • Human CCL3/MIP-1 alpha Quantikine ELISA Kit
    Name: Adam Guess
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 10/14/2016
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Protocols

View specific protocols for Human CCL3/MIP-1 alpha Quantikine ELISA Kit (DMA00):

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. For Serum & Plasma Samples Only: Add 50 µL Assay Diluent
  3.   For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  8.   Aspirate and wash 3 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. 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.

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