- Regulatory Status
- Other Names
- IL-6, GM-CSF, IL-3, IL-11, SCF, CXCL12, SDF-1
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- Product Citations
Hematopoietic stem cells (HSCs) are located in the red bone marrow, and differentiate to give rise to myeloid, erythroid, and lymphoid lineages of cells which can further differentiate into specialized blood cells. The differentiation of HSCs into specialized cells is orchestrated by a complex interplay between cytokines, growth factors, and chemokines which are considered important lineage-specific markers.
The LEGENDplex™ Human HSC Erythroid Panel (6-plex) is a bead-based multiplex assay panel, which uses fluorescence-encoded beads suitable for use on various flow cytometers. This Panel focuses on 6 key targets involved in the differentiation of erythroid progenitor cells into erythrocytes and platelets: IL-3, IL-6, IL-11, GM-CSF, SCF, and CXCL12 (SDF1). It provides higher detection sensitivity and broader dynamic range than traditional ELISA methods. This panel has been validated for use on cell culture supernatant, serum, and plasma samples.Product Details
- Kit Contents
- Setup Beads 1: FITC Beads (1 ml)
- Setup Beads 2: PE Beads (1 ml)
- Setup Beads 3: Raw Beads (2 ml)
- Capture Beads (Varies)
- Detection Antibodies (3.5 ml)
- Standard Cocktail, Lyophilized (1 vial)
- SA-PE (3.5 ml)
- Matrix A (lyophilized)
- Lyophilized Standard Reconstitution Buffer (1 ml)
- Assay Buffer (25 ml)
- Wash Buffer (25 ml)
- Plate Sealers (4 sheets)
- Data Analysis Software Dongle
- V-bottom 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 Centrifuges
- A Vacuum Filtration Unit and a Vacuum Source (if using filter plates)
- If I don't have a vacuum, how do I remove the liquid from my plate?
If you do not have a vacuum, the assay should be run in a V-bottom plate. After centrifugation using a swinging-bucket rotor with a plate adaptor, you can remove the liquid by flicking the plate quickly, dumping the contents into a sink, and patting it dry carefully on a stack of clean paper towels without losing the beads. Alternatively, you can remove the liquid by using a pipette.
- 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. A vacuum filtration unit is required to work with the filter plates. However, if you don’t have access to a vacuum manifold or if you prefer, then you can use the V-bottom plates and follow the recommended assay protocols for the type of plates you choose. All plates should be made from low binding polypropylene. Polystyrene ELISA or cell culture plates should not be used.
- 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.
- What is the shelf life of LEGENDplex™ kits?
LEGENDplex™ kits are guaranteed for 6 months from the date of receipt, but may have a shelf life of up to 2 years from the date of manufacture.
- 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). Other software may be available to analyze FCS files. Data generated using LEGENDplex™ kits can be analyzed using the freely available LEGENDplex™ data analysis software. Please check our website for the most updated versions of the software.