Human CCL4/MIP-1 beta Quantikine ELISA Kit

R&D Systems | Catalog # DMB00

R&D Systems
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Key Product Details

Assay Length

3.0 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (10 µL), Serum (150 µL), EDTA Plasma (150 µL), Heparin Plasma (150 µL), Citrate Plasma (150 µL)

Sensitivity

4 pg/mL (Cell Culture Supernates)
11 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)

Assay Range

15.6-1000 pg/mL (Cell Culture Supernates)
31.2-2000 pg/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human CCL4/MIP-1 beta Quantikine ELISA Kit

The Quantikine Human MIP-1 beta Immunoassay is a 3.0 or 4.5 hour solid phase ELISA designed to measure MIP-1 beta in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-1 beta (Act-2 variant) and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant human MIP-1 beta. Results obtained using natural human MIP-1 beta 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 for natural human MIP-1 beta.

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 beta

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 40 40 40
Mean (pg/mL) 23.3 98.8 428 31.4 118 476
Standard Deviation 1.1 3.8 17.3 3.7 9.9 28.0
CV% 4.7 3.8 4.0 11.8 8.4 5.9

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 51.3 208 932 58.8 220 949
Standard Deviation 4.6 7.5 30.2 5.7 15.6 70.5
CV% 9.0 3.6 3.2 9.7 7.1 7.4

Recovery for Human CCL4/MIP-1 beta Quantikine ELISA Kit

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 107 81-117
Citrate Plasma (n=5) 99 87-116
EDTA Plasma (n=5) 100 84-112
Heparin Plasma (n=5) 99 90-112
Serum (n=5) 99 88-113

Linearity

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

Human CCL4/MIP-1 beta ELISA Linearity

Scientific Data Images for Human CCL4/MIP-1 beta Quantikine ELISA Kit

Human CCL4/MIP-1 beta ELISA Cell Culture Supernate Standard Curve

Human CCL4/MIP-1 beta ELISA Cell Culture Supernate Standard Curve

Human CCL4/MIP-1 beta ELISA Serum/Plasma Standard Curve

Human CCL4/MIP-1 beta 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: CCL4/MIP-1 beta

CCL4/MIP-1 beta is a beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells. It attracts a variety of immune cells to sites of microbial infection as well as to other pathologic inflammation such as allergic asthma and ischemic myocardium. CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha. It signals through CCR5, and an N-terminally trimmed form additionally interacts with CCR1 and CCR2. In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor.

Alternate Names

Exodus-3, MIP-1 beta, MIP1 beta

Entrez Gene IDs

6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine); 102117861 (Cynomolgus Monkey)

Gene Symbol

CCL4

Additional CCL4/MIP-1 beta Products

Product Documents for Human CCL4/MIP-1 beta 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 CCL4/MIP-1 beta 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 CCL4/MIP-1 beta Quantikine ELISA Kit

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Protocols

View specific protocols for Human CCL4/MIP-1 beta Quantikine ELISA Kit (DMB00):

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. Add Assay Diluent
  3.   For Serum & Plasma Samples: Add 100 µL of Assay Diluent to each well.
    For Cell Culture Supernate Samples: Add 50 µL Assay Diluent to each well.
    4. Add Standard, Control, or Sample
  4.   For Serum & Plasma Samples: Add 150 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1.25 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 1.25 hours.
  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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