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FlowPlex Multi-Analyte Quantitation

Quick finder . . . Summary FlowPlex - a sandwich assay on a bead Distinguishing Up to 20 analytes by flow cytometry Advantages of FlowPlex Human FlowPlex Assays Mouse FlowPlex Assays Rat FlowPlex Assays Reagents Provided Create your own Assay Free Demo Available Download Software

FlowPlex – a bead based sandwich assay allowing simultaneous quantitation of up to 20 analytes

• Configure the assay to meet your personal requirements:
  • Choose from pre designed Multiplex Kits or single analyte Simplex kits.
  • Up to 20 analytes can be combined to create your assay of choice.
• Only 25ul of sample required for up to 20 analytes.
• Compatible with all flow cytometers.
• Free analytical software, license free. Can be loaded onto any computer.
• Free on site demonstration of the Human or Mouse Th1/Th2 FlowPlex Kits.
  • Allows the analysis of up to 16 of your own samples.
  • Call 0800 279 9113 to make a booking.

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FlowPlex - a sandwich assay on a bead

Beads are coupled with antibodies specific to the analyte to be detected. A mix of coupled beads is incubated with the samples, and analytes present in the sample bind to the antibodies coupled to the fluorescent beads. A biotin-conjugated antibody mix is added which binds to the analytes captured by the first antibodies. Streptavidin-phycoerythrin (PE) is added which binds to the biotin conjugates. Beads are differentiated by their size and distinct spectral addresses by flow cytometry.

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Distinguishing up to 20 Analytes by Flow Cytometry

Individual beads can easily be detected by any flow cytometer with one or more laser. Beads of two different sizes (5µm & 4µm) are used in the FlowPlex multiplex assay. Each of the two sets consists of a range of bead populations internally dyed with varying intensities of Starfire Red - a fluorescent dye. Starfire Red can be excited with any FACS laser and emits at 690nm, which is detected in the FL-3/FL-4 channel. The combination of the two different bead sizes and different internal dye intensities makes it possible to distinguish up to 20 bead sets in one fluorescent channel. PE, which binds to the biotin conjugate, emits at 578nm, is detected in the FL-2 channel and allows the quantification of the analyte.

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Advantages of FlowPlex

• Only 25µl of sample required to measure up to 20 analytes.
• Serum, plasma, cell culture supernatants, whole blood and other bodily fluids can all be measured.
• The dynamic range is up to 20 000 pg/ml.
• Compatible with most automated flow cytometers with an excitation capability of 488 nm.
• Pre-designed panels are available to meet most customers’ requirements.
• For maximum flexibility simplex kits can be configured to create your assay of choice.
• Use of 96 well filter plates, which are provided with the kit, reduces bench time.
• Data correlates closely with ELISA kits from most suppliers.
• Excellent sensitivity and reproducibility.
• Human standards are validated against NIBSC standards.

Software is free and easy to use

• FlowPlex Pro Software enables complete analysis of data files and calculation of sample values by comparing them against the standard curve.
• Software is compatible with PC and Macintosh computers.
• Sample results can be obtained within minutes of completing an experiment.
• A five-parameter logistic curve (5PL) is used for precise analysis of standard curves; this is the most accurate method for the estimation of asymmetric immuno-assay data.
• FlowPlex software allows measurement of standards and samples in duplicate and calculation of coefficient of variation (CV%).

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Human FlowPlex Assays

Human Multiplex Kits

Product	Cat. No.	Description	Price (£)	Price (€)
Th1/Th2 11 Plex Kit	UKB810	IFN-γ,IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12,TNF-α,TNF-β	1309	2094
Adhesion 6 Plex Kit	UKB812	sE-selectin, sICAM-1, sICAM-3, sP-selectin, sVCAM-1, sPECAM-1	744	1190
Cardiovascular 7 Plex Kit	UKB811	sCD40L, IL-6, IL-8, MCP-1, sP-selectin, t-PA, VCAM-1	862	1379
Chemokine 6 Plex Kit	UKB813	G-CSF, IL-8, MCP-1, MIG, MIP-1α,MIP-1β	744	1190

Additional Simplex assays can be combined with any of the multiplex assays detailed above to a maximum of 20 analytes. Simplex kits with identical bead populations cannot be combined click here to see the possibilities.

Human Simplex Kits

Analyte	Cat. No.	Bead Population	Sensitivity (pg/ml)	Price (£)	Price (€)
E-Selectin*	UKB8205	A9	2900	153	245
G-CSF	UKB82001	A11	6.35	153	245
ICAM-1*	UKB8201	A11	8100	153	245
ICAM-3*	UKB8218	A7	500	153	245
IFN-α	UKB82163	B3	8.06	153	245
IFN-γ	UKB8228	A4	2.2	153	245
IL-10	UKB8215	A8	1.5	153	245
IL-12	UKB8238	A2	11.6	153	245
IL-13	UKB8231	B4	4.5	153	245
IL-18	UKB8267	B5	13.1	153	245
IL-1β	UKB8224	B8	5.2	153	245
IL-2	UKB8221	A6	20.0	153	245
IL-4	UKB8225	B6	15.7	153	245
IL-5	UKB8278	B7	10.0	153	245
IL-6	UKB8213	A12	0.8	153	245
IL-8	UKB8204	A10	7.9	153	245
MCP-1	UKB8281	A7	11.0	153	245
MIG	UKB8285	B4	3.31	153	245
MIP 1-α	UKB82029	A9	6.44	153	245
MIP 1-β	UKB82030	A5	2.26	153	245
PECAM-1*	UKB8229	A5	1300	153	245
P-Selectin*	UKB8219	B4	100	153	245
sCD30	UKB8240	B3	5.1	153	245
sCD40L	UKB8239	A3	50.0	153	245
sVCAM-1*	UKB8232	B5	100	153	245
TNF-α	UKB8223	B9	17.5	153	245
TNF-β	UKB8202	B10	3.1	153	245
TNF-R (60kDa)	UKB8203	A5	100	153	245
tPA	UKB8258	A5	8.0	153	245

Human Basic Kit

The Human basic kit is required to run Simplex kits, unless Simplex kits are being combined with a Human Multiplex kit.

Product	Description	Cat. No.	Price (£)	Price (€)
Human Basic kit	For use with simplex kits: G-CSF, IFN-α,IFN-γ, IL-10, IL-12, IL-13, IL-18, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, MCP-1, MIG, MIP 1-α, MIP 1-β, PECAM-1, sCD30, sCD40L, sVCAM-1, TNF-α, TNF-β, TNF-R (60kDa), tPA	UKB8420	169	270
Human basic kit PE	For use with simplex kits: E-selectin, ICAM-1, ICAM-3, P-selectin, PECAM-1,sVCAM-1	UKB8421	169	270

If Human Simplex kits PE* are combined with other simplex kits only the basic kit UKB8420 is required.

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Mouse FlowPlex Assays

Mouse Multiplex Kits

Product	Cat. No.	Description	Price (£)	Price (€)
Th1/Th2 10 Plex Kit	UKB820	GM-CSF, IFN-γ, IL-1α, IL-2,IL-4, IL-5, IL-6, IL-10, IL-17,TNF-α	1237	1979
Chemokine 6 Plex Kit	UKB821	GM-CSF, MCP-1, MCP-3, MIP-1α, MIP-1β, RANTES	744	1190

Additional simplex assays can be combined with any of the multiplex assays detailed above to a maximum of 20 analytes.

Mouse Simplex Kits

Analyte	Cat. No.	Bead Population	Sensitivity (pg/ml)	Price (£)	Price (€)
GM-CSF	UKB8612	B8	15	153	245
IFN-γ	UKB8606	B6	4.0	153	245
IL-1α	UKB8611	A4	19.7	153	245
IL-2	UKB8601	A6	7.5	153	245
IL-4	UKB8613	B9	7.3	153	245
IL-5	UKB8610	A8	4.1	153	245
IL-6	UKB8603	A10	0.9	153	245
IL-10	UKB8614	A12	13.4	153	245
IL-17	UKB8601	B10	2.8	153	245
MCP-1	UKB86005	A9	50	153	245
MCP-3	UKB86006	A7	9	153	245
MIP 1-α	UKB86013	A11	17	153	245
MIP 1-β	UKB860014	B2	82	153	245
RANTES	UKB86009	B4	16	153	245
TNF-α	UKB8607	B7	8.6	153	245

Mouse Basic Kit

The Mouse basic kit is required to run Simplex kits, unless Simplex kits are being combined with a Mouse Multiplex kit.

Product	Description	Cat. No.	Price (£)	Price (€)
Mouse Basic kit	For use with simplex kits:	UKB8440	169	270

Rat FlowPlex Assays

Product	Cat. No.	Description	Price (£)	Price (€)
Th1/Th2 6 Plex Kit	UKB725	GM-CSF, IFN-γ, IL-1α, IL-4, MCP-1, TNF-α	744	1190

Rat Simplex Kit

Analyte	Cat. No.	Bead Population	Sensitivity (pg/ml)	Price (£)	Price (€)
GM-CSF	UKB633	B3	5.5	153	245
IL-1α	UKB627	A1	2.2	153	245
IL-4	UKB628	B4	1.6	153	245
MCP-1	UKB631	A2	1.2	153	245
TNF-α	UKB622	A5	5.1	153	245

Rat Basic Kit

The Rat basic kit is required to run Simplex kits, unless Simplex kits are being combined with a Multiplex kit.

Product	Description	Cat. No.	Price (£)	Price (€)
Rat Basic kit	For use with simplex kits:	UKB8440	169	270

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Reagents Provided for FlowPlex assays

Multiplex Kits	Simplex Kits	Basic Kits
Fluorescent beads coated with	Fluorescent beads coated with	Streptavidin-PE
corresponding antibody	corresponding antibody	96-well Filter plate + plate covers
Reference Standard	Reference Standard	Assay and dilution buffer
Detection antibody	Detection antibody	Setup beads
Streptavidin-PE		CD-Rom (software)
96-well Filter plate + plate covers
Assay and dilution buffer
Setup beads
CD-Rom (software)

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Create your own Assay

FlowPlex multiplex and simplex kits can be combined to create the assay of your choice, allowing the simultaneous detection of up to 20 analytes. Simplex kits with identical bead populations cannot be combined.

» click here to see the possibilities

Free Demo Available

If you are interested in an on site demonstration of FlowPlex kits, please let us know, call 0800 279 9113 or +44 (0) 1296 715459 and register your interest.

The day would include a visit from our technical expert for the day to demonstrate to you with your samples on your machine how easy it is to generate data using FlowPlex.

Software

Do you want several copies of the FlowPlex data handling software? Allowing you to do your data analysis on a machine of your choice! We can send you the software on a CD or you may download it here:

Download FlowPlex Set Up V2.2
Please note: The file is an Windows executable "set-up" programme. It is a large file (21mB) so will take time to download.
Save it to your computer ("right click" and "save target as") and then double click the downloaded file to start the install.

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