Human CCL4/MIP-1 beta XL Luminex® Performance Assay

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MIP-1 beta XL Cytokine Luminex Panel Standard Curves
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Human CCL4/MIP-1 beta XL Luminex® Performance Assay Summary

Panel Luminex Performance Human XL Cytokine Panel
Assay Type Magnetic bead-based multiplex assay for the Luminex® platform
Format 1 x 96-well microplate and Magnetic antibody-coated beads
Species Human
Analytes Detected Please see analyte list in assay customization tool below.
Performance Validation Our Luminex High Performance Assays undergo our most extensive validation testing. All Luminex High Performance Assay Panels are validated for use with serum and plasma samples. Some High Performance Assays are additionally validated for cell culture supernatants, milk, saliva, or urine, as indicated on the individual product data sheets. Luminex High Performance Assays are tested for sensitivity (three-quarters the low standard), intra-assay precision, inter-assay precision, and to ensure assay linearity for validated sample types. Recovery values for individual samples in validated sample types are also tested. Validation data for each analyte can be found in the product datasheet.

View Luminex High Performance Human XL validation data.

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Assays

Below are kits that are available based on your selections. These selections may be further customized before adding to the cart or emailing the configuration to your purchasing department or local distributor to place your order. Selected analytes may be included in more than one High Performance Assay. All assay options are displayed.

If you run into bead region incompatibilities, send your Luminex code to our team and find out more.

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Not finding your analyte of interest? View our complete Luminex offering in the full version of our Luminex Assay Customization Tool. We also offer Custom Luminex Services.

Product Features

Magnetic Luminex High Performance Assays are flexible bead-based multiplex assays. They allow up to 50 user-defined target analytes to be simultaneously profiled using cell culture supernates, serum, or plasma samples. Some panels are also validated with human milk, saliva, and/or urine samples.

  • Simultaneously profile Human CCL4/MIP-1 beta with up to analytes of your choice in one sample
  • Magnetic format allows for easier wash steps
  • Undergoes similar validation testing as Quantikine® ELISA Kits
  • Precise and Specific
  • Requires a small sample volume (<50 µL)
  • Run your assay in just 3-5 hours
  • Mass-calibrated standards for consistent results with every new lot of material
  • Enough reagents for one 96-well plate
Base Kit Components
  • Premixed cocktail of biotinylated detection antibody
  • Standard Cocktail(s)
  • Bead Diluent
  • Biotin Antibody Diluent
  • Calibrator Diluent
  • Wash Buffer
  • Streptavidin-PE
  • One flat-bottom 96-well Microplate
  • Foil Plate Sealers (4)
  • Mixing Bottle
  • Standard Value Card**
  • High and low controls with Control Mean Value Card

**A standard curve must be generated each time an assay is run, utilizing values from the Standard Value Card included in the Base Kit.

Bead Set Contents
  • User-mix antibody-coated Magnetic beads in individual vials

Recommended Sample Types: Cell culture supernates, serum, EDTA plasma, and heparin plasma

Components: Human CCL4/MIP-1β Magnetic Microparticles (Part 898824) is supplied as a 100X concentrated stock (0.075 mL) with preservatives.

Other Supplies Required: Magnetic Luminex® Performance Assay Human XL Discovery Base Kit (R&D Systems®, Catalog # LUXLM000).

Sensitivity: The Minimum Detectable Dose (MDD) was determined by adding two standard deviations to the mean MFI of twenty zero standard replicates and calculating the corresponding concentration.
Six assays were evaluated, and the MDD of human MIP-1β ranged from 5.23-72.9 pg/mL. The mean MDD was 18.4 pg/mL.

Correlation: This assay has been correlated to the Quantikine® ELISA Kit.

Specificity: This assay recognizes natural and recombinant human MIP-1β.

Bead Region: 21
 

Precision

Intra-assay Precision (precision within an assay) - Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (precision between assays) - Two samples of known concentration were tested in thirty separate assays to assess precision between assays.

  Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 20 20 30 30
Mean (pg/mL) 509 3066 525 3,187
Standard deviation 7.82 55.7 51.5 191
CV (%) 53.6 81.6 81.3 2.1
 

Recovery

Samples containing and/or spiked with human MIP-1 beta were evaluated for recovery.

Sample Type Average % Recovery Range (%)
Cell culture supernate 134 121-163
Serum 94.9 64.8-121
EDTA plasma 100 72.9-125
Heparin plasma 95.4 67.3-120
Plaelet-poor EDTA 106 78.2-127
Platelet-poor Heparin 100 73.2-115
 

Linearity

Samples containing and/or spiked with human MIP-1 beta were serially diluted to evaluate assay linearity.

  Cell culture supernate Serum EDTA plasma Heparin plasma Platelet-poor EDTA Platelet-poor Heparin
1:2 Average % of Expected 82.9 108 108 117 105 110
Range (%) 82.0-83.9 99.7-123 99.1-117 105-132 95.8-115 102-116
1:4 Average % of Expected 75.2 106 105 114 103 111  
Range (%) 72.6-77.6 90.4-121 91.4-120 100-126 87.6-115 93.2-126
1:8 Average % of Expected 76.3 112 111 120 110 115
Range (%) 73.9-79.8 93.0-126 93.4-129 103-136 91.7-123 99.8-128
1:16 Average % of Expected 76.6 109 109 116 109 111
Range (%) 73.4-80.3 92.3-127 91.4-126 99.5-132 91.1-126 99.9-124
Which Luminex Assay is right for me?

Assays for the Luminex platform are offered as High Performance Assays or Assays. The Luminex High Performance Assays are fully validated panels with a focused selection of analytes. They are available in We Mix, You Mix, and Predetermined formats. The Luminex Assays allows for the maximum number of analytes in a multiplex and is supplied as a premixed kit. View a table comparing the features and benefits of these bead-based multiplex assays.

More Information About Magnetic Bead-based Multiplex Kits For The Luminex® Platform

Magnetic Luminex High Performance Assays are designed for use with the Luminex MAGPIX CCD Imager. Alternatively, kits can be used with the Luminex 100/200™ or FLEXMAP 3D, dual laser, flow-based sorting and detection platforms.

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. One laser is bead-specific and determines which analyte is being detected. A magnet in the analyzer captures and holds the superparaMagnetic microparticles in a monolayer. Two spectrally distinct Light Emitting Diodes (LEDs) illuminate the beads. One LED identifies the analyte that is being detected and the second LED determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of analyte bound. Each well is imaged with a CCD camera. Kits can also be used with Luminex 100/200 or a Bio-Rad Bio-Plex dual laser, flow-based systems.

Scientific Data

MIP-1 beta XL Cytokine Luminex Panel Standard Curves

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.

Entrez Gene IDs
6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine); 102117861 (Cynomolgus Monkey)
Alternate Names
ACT2; ACT-2; AT744.1; C-C motif chemokine 4; CCL4; chemokine (C-C motif) ligand 4; Exodus-3; G-26 T-lymphocyte-secreted protein; G-26; HC21; LAG1; LAG-1; Lymphocyte activation gene 1 protein; Macrophage inflammatory protein 1-beta; MIP1 beta; MIP-1 beta; MIP1B; MIP1B1; MIP-1-beta; MIP-1-beta(1-69); PAT 744; Protein H400; SCYA2; SCYA4; secreted protein G-26; SIS-gamma; small inducible cytokine A4 (homologous to mouse Mip-1b); Small-inducible cytokine A4; T-cell activation protein 2

Product Datasheets

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Product Specific Notices
Luminex® is a registered trademark of Luminex Corporation.

Citation for Human CCL4/MIP-1 beta XL Luminex® Performance Assay

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Multiple cytokine analyses of aqueous humor from the patients with retinitis pigmentosa
    Authors: B Lu, H Yin, Q Tang, W Wang, C Luo, X Chen, X Zhang, K Lai, J Xu, X Chen, K Yao
    Cytokine, 2019-12-03;127(0):154943.
    Species: Human
    Sample Types: Aqueous Humor

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