- Other Names
- Human Bone Formation Panel (4-plex)
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Bone metabolism is a complex equilibrium between bone formation and resorption regulated through numerous hormones, steroids, growth factors, and cytokines. Bone formation is mediated through specialized cells called osteoblasts, while resorption is carried out via osteoclasts. Bone metabolism is a constant, life-long process that exists in balance. Disruptions to the homeostasis between formation and resorption results in diseases such as: osteoporosis, osteopetrosis, Paget’s disease, rickets, and renal osteodystrophy.
The LEGENDplex™ Human Bone Formation Panel (4-plex) is a bead-based multiplex assay panel, using fluorescence-encoded beads suitable for use on various flow cytometers. It allows for simultaneous quantification of 4 key targets involved in bone formation including OPG, ALPL, Leptin, and BMP-2. This assay panel provides higher detection sensitivity and broader dynamic range than traditional ELISA methods. The panel has been validated for use on cell culture supernatant, serum, and plasma samples.
- Kit Contents
- Setup Beads 1: FITC Beads (1 ml)
- Setup Beads 2: PE Beads (1 ml)
- Setup Beads 3: Raw Beads (1 ml)
- Capture Beads (varies)
- Detection Antibodies (3.5 ml)
- Standard Cocktail, Lyophilized (1 vial)
- SA-PE (3.5 ml)
- Matrix C, Lyophilized (1 vial)
- Reconstitution Buffer (1 ml)
- Assay Buffer (25 ml)
- Wash Buffer (25 ml)
- Plate Sealers (4 sheets)
- Data Analysis Software Dongle
- Filter Plate
Learn more about LEGENDplex™ at biolegend.com/legendplex
Download the LEGENDplex™ software here.
- Materials Not Included
- Flow Cytometer
- Pipettes and Tips
- Reagent Reservoirs for Multichannel Pipettes
- Polypropylene Microfuge Tubes
- Vortex Mixer
- Aluminum foil
- Absorbent pads or paper towels
- Plate Shaker
- Tabletop centrifuge
- A Vacuum Filtration Unit and a Vacuum Source (if using filter plates)
- After I finish the staining process, how long can I wait before reading my LEGENDplex™ samples?
The samples can be kept overnight at 4°C while being protected from exposure to light and be read the next day. There may be a decrease in signal, but overall, the assay results should not be affected. Storing the samples for extended periods of time is not recommended, as it could lead to further reductions in signal.
- If I don't have a vacuum, how do I remove the liquid from my plate?
Do NOT invert the plate to dump the liquid. The beads will not adhere to the plate. After centrifugation using a swing bucket rotor with a plate adaptor, you can remove the liquid by using a pipette.
- Is special software required for data analysis?
Typically flow cytometers generate output files in FCS format (e.g. FCS 2.0, 3.0, or 3.1) and in some cases in list mode file format (LMD). FCS files can be analyzed by different software. Data generated from using LEGENDplex™ kits can be analyzed using the LEGENDplex™ data analysis software freely available on the BioLegend website (biolegend.com/legendplex/software). A free of charge software license dongle is currently provided in the kit. Please check our website for the most updated versions of the software.
- Should I perform the assay with the filter plates or with V-bottom plates?
Filter plates or V-bottom plates have been included in some kits for your convenience. Performing the assay with filter plates is generally recommended. A vacuum filtration unit is required to work with the filter plates. If however, you don’t have access to a vacuum manifold, then you can use the V-bottom plates or micro-FACS tubes as described in the manual and follow the recommended assay protocols for the type of plates or tubes you choose. All tubes and plates should be made from low binding polypropylene. Polystyrene ELISA or cell culture plates should not be used.
- What is the shelf life of LEGENDplex™ kits?
LEGENDplex™ kits are valid for 6 months from the date of receipt.