GeniePlex MagPro Human Endothelial Activation 4-Plex (MPES0040)

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SKU:
MPES0040
Product Type:
Multiplex
Reactivity:
Human
Analytes:
sICAM-1, sVCAM-1, MMP-9, and VEGF
Frequently bought together:

Description

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GeniePlex MagPro Human Endothelial Activation 4-Plex is a magnetic bead-based multiplex immunoassay kit for the simultaneous quantitative detection of 4 analytes from a single sample using flow cytometry. Antibody-conjugated beads with distinct fluorescence intensities capture target antigens, which are then detected via biotinylated antibodies and streptavidin-PE to produce a fluorescent signal proportional to each analyte concentration.

Product Name:GeniePlex MagPro Human Endothelial Activation 4-Plex (96 Tests)
Product Code:MPES0040
Reactivity:Human
Analytes (4):sICAM-1, sVCAM-1, MMP-9, and VEGF
Range of Linearity:sICAM-1: 40–10,000 pg/mL, sVCAM-1: 40–10,000 pg/mL *, MMP-9: 10–5,000 pg/mL, VEGF: 10–5,000 pg/mL (* Estimated — may vary in the final product)
Sensitivity (LODs):sICAM-1: < 20 pg/mL, sVCAM-1: < 20 pg/mL *, MMP-9: < 5 pg/mL, VEGF: < 5 pg/mL (* Estimated — may vary in the final product)
Sample Types:Serum, EDTA plasma, cell culture supernatants, and other biological fluids
Assay Type:Magnetic bead-based multiplex immunoassay
Detection Method:Flow cytometry (PE, APC, and APC/Cy7 channels)
Sample Volume:50 µL/well
Assay Time:~2.5 hours
Kit Size:96 tests
ComponentQuantity (96T)Storage
Premixed Antibody-Conjugated Beads2.4 mL × 22–8 °C, protected from light
Biotinylated Detection Antibodies4.8 mL × 22–8 °C
SA-PE (ready to use)4.8 mL × 22–8 °C, protected from light
Lyophilized Standard2 vials2–8 °C
Assay Buffer5 mL × 12–8 °C
Wash Buffer30 mL × 22–8 °C
Plate Sealing Film5 pieces—
Manual1 copy—

Storage & Stability

ConditionDetails
Unopened kit2–8 °C, protected from light — stable for 12 months
Opened kit2–8 °C, protected from light — use within 30 days
Reconstituted standard2–8 °C — use within 24 hours

Materials Required but Not Supplied

  • U-bottom 96-well transparent plates
  • Vortex mixer
  • Incubator / microplate shaker
  • Magnetic separator for 96-well plates
  • Flow cytometer with PE, APC, and APC/Cy7 detection channels
Range of Linearity:sICAM-1: 40–10,000 pg/mL, sVCAM-1: 40–10,000 pg/mL *, MMP-9: 10–5,000 pg/mL, VEGF: 10–5,000 pg/mL (* Estimated — may vary in the final product)
Limit of Blank (LoB):≤ 8 pg/mL for all analytes
Recovery:70–120%
Intra-assay CV:< 15%
Inter-assay CV:< 15%
Cross-reactivity:No significant cross-reactivity among panel analytes

*Note: The below is a summary protocol. Protocols may vary by lot. Always follow the instructions included with your kit.

Standard Preparation

  1. Label eight 0.6 mL microcentrifuge tubes 0–7. Add 150 µL Assay Buffer to tubes 0–6. Leave tube 7 empty.
  2. Reconstitute the lyophilized standard: centrifuge briefly (500 × g, 10 s), add 500 µL Assay Buffer, stand 5 min, mix gently until dissolved. Transfer to tube 7 (highest standard).
  3. Serial dilution: transfer 50 µL from tube 7 → tube 6 (1:3.2), then 100 µL from tube 6 → 5 → 4 → 3 → 2 → 1, mixing at each step. Tube 0 = zero standard (buffer only).

Assay Procedure

StepProtocol
1. Bead & Sample IncubationAdd 50 µL Premixed Antibody-Conjugated Beads (vortex ≥ 15 s) and 50 µL sample or standard to each well. Seal plate. Incubate on shaker at RT, 600 rpm, protected from light, 1 hour.
2. Detection AntibodyPlace plate on magnetic separator for 1 min, remove supernatant. Add 100 µL Biotinylated Detection Antibodies. Seal and incubate at RT, 600 rpm, protected from light, 1 hour.
3. SA-PE & WashMagnetic separate 1 min, remove supernatant. Wash once with 200 µL Wash Buffer. Add 100 µL SA-PE. Seal and incubate at RT, 600 rpm, protected from light, 30 min. Wash twice with 200 µL Wash Buffer.
4. AcquisitionResuspend beads in 200 µL Wash Buffer. Acquire on flow cytometer using PE, APC, and APC/Cy7 channels.

Data Analysis

  1. Collect at least 200 beads per analyte gate (minimum 800 total beads for this 4-plex panel).
  2. Calculate median fluorescence intensity (MFI) for standards and samples. Subtract blank MFI.
  3. Plot standard curve (concentration vs. MFI, log-log) and fit with a 4-parameter logistic (4-PL) model.
  4. Determine sample concentrations from the standard curve. If MFI exceeds the upper limit, dilute and re-measure.
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