Human CD40 Ligand/TNFSF5 XL Luminex® Performance Assay

Name: Human CD40 Ligand/TNFSF5 XL Luminex® Performance Assay
Brand: R&D Systems
SKU: LUXLM617
Price: 189 USD

Catalog #: LUXLM617 Datasheet / COA / SDS

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CD40 Ligand XL Cytokine Luminex Panel Standard Curves

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Human CD40 Ligand/TNFSF5 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.

Make your selections below to configure your assay

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.

How would you like these analytes mixed?

Premixed

User Mixed

Please select at least one mixing option to see the kits.

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Do not split

Split based on serum

Split based on plasma

Split based on cell culture supernates

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 CD40 Ligand/TNFSF5 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, platelet-poor plasma

Components: Human CD40 Ligand/TNFSF5 Magnetic Microparticles (Part 898852) 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 CD40 Ligand ranged from 49.5-379 pg/mL. The mean MDD was 110 pg/mL.

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

Bead Region: 25

Precision

	Intra-Assay Precision		Inter-Assay 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.
Sample	1	2	1	2
n	20	20	30	30
Mean (pg/mL)	5323	52,667	5450	52,704
Standard deviation	154	2455	766	3993
CV (%)	89.3	66.1	14.0	57.6

Recovery

Sample Type	Average % Recovery	Range (%)
Samples containing and/or spiked with human CD40 Ligand were evaluated for recovery.
Cell culture supernate	114	105-126
Serum	75	61-102
EDTA plasma	87	66-111
Heparin plasma	86	70-101
Plaelet-poor EDTA	78	56-94
Platelet-poor Heparin	79	60-101

Linearity

		Cell culture supernate	Serum	EDTA plasma	Heparin plasma	Platelet-poor EDTA	Platelet-poor Heparin
Samples containing and/or spiked with human CD40 Ligand were serially diluted to evaluate assay linearity.
1:2	Average % of Expected	98	125	120	114	124	120
1:2	Range (%)	94-103	119-133	112-129	101-130	115-138	114-128
1:4	Average % of Expected	93	128	118	110	134	130
1:4	Range (%)	91-96	122-136	111-125	98-124	116-151	116-157
1:8	Average % of Expected	95	137	124	114	149	131
1:8	Range (%)	88-100	128-143	113-141	98-128	123-172	125-139
1:16	Average % of Expected	97	131	118	106	143	123
1:16	Range (%)	94-100	123-141	108-139	97-121	121-173	116-130

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

N/A

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CD40 Ligand XL Cytokine Luminex Panel Standard Curves

Background: CD40 Ligand/TNFSF5

CD40 Ligand, also known as TNFSF5, CD154, TRAP, and gp39, is a trimeric protein that is expressed in transmembrane and soluble forms. It is expressed by many immune cell types as well as by endothelial and smooth muscle cells of the vasculature. It binds to CD40 on antigen presenting cells, platelets, and endothelial and epithelial cells, leading to enhanced B cell activation and T cell dependent humoral immune responses. CD40 Ligand dysregulation contributes to immune pathology in AIDS, atherosclerosis, atherothrombosis, and restenosis.

Entrez Gene IDs

959 (Human); 21947 (Mouse); 84349 (Rat)

Alternate Names

CD154 antigen; CD154; CD40 antigen ligand; CD40 Ligand; CD40L; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5