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Invitrogen™ CD80 Mouse ProcartaPlex™ Simplex Kit

Catalog No. 502557329
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The Mouse CD80 Simplex ProcartaPlex Kit measures CD65 protein and is designed to be combinable with other Simplex kits so that you can create your own multiplex panel that utilizes Luminex xMAP technology for protein detection/quantitation.

The Mouse CD80 Simplex ProcartaPlex Kit measures CD65 protein and is designed to be combinable with other Simplex kits so that you can create your own multiplex panel that utilizes Luminex xMAP technology for protein detection/quantitation. When combining multiple Simplex kits (i.e., when you are not using a pre-configured Multiplex Panel), only one buffer kit (sold separately) is needed for each assay plate regardless of plex size.

About ProcartaPlex Assays for the Luminex Platform

ProcartaPlex immunoassays are based on the principles of a sandwich ELISA, using two highly specific antibodies binding to different epitopes of one protein to quantitate all protein targets simultaneously using a Luminex instrument. ProcartaPlex multiplex assays require as little as 25 μL of plasma or serum, or 50 μL of cell culture supernatant, and just four hours to obtain analyzed results.

  • More results per sample—measure multiple protein targets simultaneously in a single 25–50 μL sample
  • Well-established Luminex technology—highly referenced multiplexing platform for protein detection and quantitation

ProcartaPlex assays utilize Luminex xMAP (multianalyte profiling) technology for the simultaneous detection and quantitation of up to 80 protein targets in a single 25–50 μL sample from plasma, serum, cell culture supernatants, and other bodily fluids.

The Luminex beads in the ProcartaPlex assay are internally dyed with precise proportions of red and infrared fluorophores to create spectrally unique signatures that can be identified by the Luminex xMAP detection systems (e.g. Luminex 200, FLEXMAP 3D, and MAGPIX). Similar to a sandwich ELISA, the ProcartaPlex assay uses matched antibody pairs to identify the protein of interest. In a multiplexed assay, each spectrally unique bead is labeled with antibodies specific for a single target protein, and bound proteins are identified with biotinylated antibodies and streptavidin–R-phycoerythrin (RPE). The conjugation of protein-specific antibodies to a distinct bead allows for analysis of multiple targets in a single well.

The most significant difference between a ProcartaPlex assay and ELISA is that the capture antibody in the ProcartaPlex assay is conjugated to a bead and not adsorbed to the microplate well, so the ProcartaPlex assay reagents are free-floating in the solution. For detection, the Luminex 200 instrument, for example, contains two lasers, one to distinguish the spectral signature of each bead and the second to quantify the amount of RPE fluorescence, which is proportional to the amount of protein present in the sample. ProcartaPlex multiplex assays can profile more target proteins using significantly less sample in the same time that it takes to perform a traditional sandwich ELISA.

ProcartaPlex multiplex panels are available in multiple formats across six species (human, mouse, rat, nonhuman primate, porcine, and canine).

TRUSTED_SUSTAINABILITY

Specifications

Product Type Simplex Kit
Quantity 96 Tests
Species Mouse
Combinability Combinable
Bead Type CD80 [15]
Assay Sensitivity As determined for Lot 1: 7.8 pg/mL
Assay Range As determined for Lot 1: 72 - 296,200 pg/mL
Format Simplex Kit
For Use With (Equipment) Luminex™ Instruments
Gene CD80 antigen
Gene Alias B71,Cd28l,Ly-53,Ly53,MIC17,TSA1
Gene ID (Entrez) 12519
Gene Symbol CD80
Product Line ProcartaPlex
Protein T-lymphocyte activation antigen CD80
Protein Subtype B7
Sample Type Serum, Plasma, Cell Culture Supernatants
Sample Volume Serum, Plasma: 25 μL, CCS: 50 μL
UniProt ID Q00609
Content And Storage

• 1 vial Capture Beads (50X)
• 1 vial biotinylated Detection Antibody (50X)
• 2 vials Mouse Standard Mix F (lyophilized)

Store at 2°C to 8°C.

Shipping Condition Wet Ice
Research Area Immunology, Oncology
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Do you offer a Mac-compatible version of the ProcartaPlex Analyst Software?

Unfortunately, we do not currently offer a Mac-compatible version of the ProcartaPlex Analyst Software. ProcartaPlex Analyst Software is compatible with PC, or xPONENT Software is available on Luminex instruments to analyze your data.

Do you offer FlowCytomix assays?

Sorry, the FlowCytomix product line has been discontinued. However, we offer a new antibody and magnetic bead-based detection system, i.e., ProcartaPlex assays for multiplex protein quantitation using the Luminex instrument platform.

How long does it take to acquire ProcartaPlex assay data on the different Luminex instruments?

With everything working smoothly, here is the approximate read time for one plate on different Luminex instruments:

- FLEXMAP 3D Systems and INTELLIFLEX Systems will read 96 well plates in ~20 min, and 384 well plates in ~75 min.
- Luminex 100/200 Systems will read 96 well plates in ~40 min.
- MAGPIX System will read 96 well plates in ~50-60 min. Please note that these are estimates for the read times of the plates only, and do not include time for calibration or set-up of the instrument.

Is the sensitivity value for a Multiplex ProcartaPlex assay different when compared to the sensitivity for those same analytes assayed as Simplex assays?

In general, sensitivity between Simplex and Multiplex ProcartaPlex assay kits is comparable.

Why does the ProcartaPlex ULOQ (Upper Limit of Quantitation) and LLOQ (Lower Limit of Quantitation) change between different lots of standards?

We supply the standard protein in a lyophilized form. Lyophilization is a dehydration process that preserves the perishable standard. We have demonstrated that often, the amount of starting protein prior to lyophilization and that of the final product after lyophilization varies (this could be different from analyte to analyte). To ensure that we deliver the most quantitative and reproducible product, we always calibrate the standard after lyophilization and therefore, the concentration range can vary from lot to lot of the standard.

Can the ProcartaPlex beads be frozen?

No. Storage of the beads at temperatures below 0 degrees C will damage the beads and render them unusable.

What is the minimum bead count used for statistical significance/analysis in ProcartaPlex assays?

We validate and release our ProcartaPlex assays with a minimum bead count of 50. In our experience, the assays also work with a bead count of 20 though we have not validated them at this bead count.

Is ProcartaPlex assay sensitivity information based on data using the 2 hr or overnight incubation with samples?

The sensitivity information is based on the 2 hr incubation with samples.

How can I minimize inter-assay variation for ProcartaPlex assays?

To minimize inter-assay coefficient of variation between two independent assay plates, it is very important that the same operator performs the assays. Also, we recommend that you follow the incubation times exactly as they are stated in the manual in order to get consistent results. Other tips for minimizing inter-assay coefficient of variation include performing a standard curve in duplicate on each plate and using the same kit lot.

I am trying to find out whether the medium/serum used to culture my cells will interfere with the ProcartaPlex assay. Which cell culture media have you tested in-house with ProcartaPlex assays?

In-house, we test ProcartaPlex assays with RPMI + 0.5% FCS. You would have to empirically determine if your specific medium will interfere with the assay.

Note: Our customers have successfully used media containing up to 5% FBS with ProcartaPlex assays. As bovine serum contains TGF beta, the use of FBS or FCS should be avoided with TGF beta ProcartaPlex assays.

I am a Bio-Plex user. How can I generate and export CSV raw data to be analyzed using the ProcartaPlex Analysis App?

Open the Bio-Plex Manager and click on “File”, then “Document Export Properties”, then “Output CSV Format” and set “Analytes Labels” to “Name (Region)”. Choose the desired file “Destination”, and click “OK”. Click “Document Export”, then “Export”, and import the generated .CSV file into ProcartaPlex Analysis App on Thermo Fisher Connect: https://apps.thermofisher.com/apps/procartaplex

How do I process tear samples for use in a Luminex assay?

We have not specifically tested ProcartaPlex assays with tear samples in-house. However, here is a reference from the literature describing the use of ProcartaPlex assays with tear samples (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161422/pdf/pone.0107370.pdf).

How do I process breast milk samples for use in a Luminex assay?

We have tested breast milk samples in-house using the serum ProcartaPlex protocol and found no general interference with the breast milk matrix. Please note that we have only tested a few analytes (TNFa, IL-10, TNFR), however, there is no obvious reason why other analytes should not work as well.

What is the size of beads used in the ProcartaPlex assays?

The size of the beads in our ProcartaPlex assays is 6.5 µm.


For Research Use Only. Not for use in diagnostic procedures.

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