Luminex 100 IS Developer Workbench Guide Version 2.3 Manual de usuario Pagina 59
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PN 89-00002-00-084 Rev. B 53
xMAP Technology Protein Coupling Protocol
Preparation 1. Allow all reagents to warm to room temperature.
2. Using an analytical balance, weigh approximately 10 mg of
Sulfo-NHS into a tube. Repeat for EDC.
Procedure
xMAP Microsphere
Activation
1. Vortex and sonicate the stock microspheres for 20 seconds.
2. Transfer 5.0 × 10
6
of the stock microspheres to a USA Scientific
microfuge tube.
3. Centrifuge the stock microspheres at ≥ 8,000 × g for 1 to 2
minutes.
4. Aspirate the supernatant and resuspend the pelleted microspheres
in 100 µL dH
2
O by sonication for approximately 20 seconds.
5. Centrifuge the stock microspheres at ≥ 8,000 × g for 1 to 2
minutes.
6. Aspirate the supernatant. Resuspend the washed microspheres in
80 µL of Activation Buffer. Vortex and sonicate for
approximately 20 seconds.
7. Add 10 µL of 50 mg/mL Sulfo-NHS (diluted in Activation
Buffer or dH
2
O) to the microspheres and mix gently by vortex.
8. Add 10 µL of 50 mg/ML EDC (diluted in Activation Buffer or
dH
2
O) to the microspheres and mix gently by vortex.
9. Incubate for 20 minutes at room temperature with gentle mixing
by vortex at 10 minute intervals.
10. Centrifuge the activated microspheres at ≥ 8,000 × g for 1 to 2
minutes.
11. Aspirate the supernatant and resuspend the microspheres in 250
µL of Coupling Buffer by vortex and sonication for
approximately 20 seconds (see Technical note 5).
12. Centrifuge the microspheres for 1 to 2 minutes at ≥ 8,000 × g.
13. Repeat steps 11 and 12 for a total of two washes with Coupling
Buffer.
14. Resuspend the activated and washed microspheres in 100 µL of
Coupling Buffer. Vortex and sonicate for approximately 20
seconds.
- Luminex 100 1
- IS Developer 1
- Workbench Guide Version 2.3 1
- PN 89-00002-00-084 Rev. B 2
- August, 2005 2
- Contents 3
- A xMAP Protocols 49 4
- Version 1.7 61 5
- C Customize Splash Screen 63 5
- Index 65 5
- Technology 6
- Developer Workbench Guide 7
- General Guidelines 8
- Instructions 9
- Instrument Calibrators 12
- Developer 13
- Workbench 13
- Software 13
- Procedures 14
- Dialog Box Buttons 15
- Create New Product 15
- 10 PN 89-00002-00-084 Rev. B 16
- PN 89-00002-00-084 Rev. B 11 17
- 12 PN 89-00002-00-084 Rev. B 18
- PN 89-00002-00-084 Rev. B 13 19
- 14 PN 89-00002-00-084 Rev. B 20
- Quantitative Assay 21
- Template 21
- 16 PN 89-00002-00-084 Rev. B 22
- PN 89-00002-00-084 Rev. B 17 23
- Note: You can use the same 23
- 18 PN 89-00002-00-084 Rev. B 24
- Figure 7. Select Bead IDs 24
- PN 89-00002-00-084 Rev. B 19 25
- Qualitative Assay 27
- 22 PN 89-00002-00-084 Rev. B 28
- PN 89-00002-00-084 Rev. B 23 29
- 24 PN 89-00002-00-084 Rev. B 30
- PN 89-00002-00-084 Rev. B 25 31
- Figure 13. Select Bead IDs 31
- 26 PN 89-00002-00-084 Rev. B 32
- PN 89-00002-00-084 Rev. B 27 33
- PN 89-00002-00-084 Rev. B 29 35
- 30 PN 89-00002-00-084 Rev. B 36
- PN 89-00002-00-084 Rev. B 31 37
- 32 PN 89-00002-00-084 Rev. B 38
- Figure 21. Select Bead IDs 38
- PN 89-00002-00-084 Rev. B 33 39
- 34 PN 89-00002-00-084 Rev. B 40
- Maintenance 41
- Edit an Unused 42
- Edit a Used 43
- To edit a used template: 43
- Delete an Unused 44
- Delete a Used 45
- To delete a used template: 45
- Print Template 46
- Information 46
- PN 89-00002-00-084 Rev. B 41 47
- Technology 48
- To import a template: 49
- To export a template: 50
- About Developer 51
- Luminex Technical 53
- Support Website 53
- Handling 55
- Enumeration of xMAP 56
- Microsphere Suspensions 56
- PN 89-00002-00-084 Rev. B 51 57
- Protein Coupling Protocol 58
- Procedure 59
- Coupling 60
- Blocking, and 60
- PN 89-00002-00-084 Rev. B 55 61
- Equipment •Vortex 62
- PN 89-00002-00-084 Rev. B 57 63
- Enumerate the Coupled 64
- Microspheres 64
- LumAvidin Binding Protocol 65
- 60 PN 89-00002-00-084 Rev. B 66
- Version 1.7 67
- 62 PN 89-00002-00-084 Rev. B 68
- Customize Splash Screen 69
- 64 PN 89-00002-00-084 Rev. B 70
- PN 89-00002-00-084 Rev. B 65 71
- 66 PN 89-00002-00-084 Rev. B 72
- PN 89-00002-00-084 Rev. B 67 73
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