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Luminex 100 IS Developer Workbench Guide Version 2.3 xMAP Technology
54 PN 89-00002-00-084 Rev. B
Coupling,
Blocking, and
Storage
1. Add the protein preparation to the resuspended microspheres.
2. Bring total volume to 500 µL with Coupling Buffer. Vortex.
3. Rotate the mixture for 2 hours in the dark at room temperature.
4. Centrifuge the microspheres 1 to 2 minutes at ≥ 8,000 × g.
Aspirate the supernatant.
5. If you plan to use the coupled microspheres on the day of
coupling, resuspend the microspheres in 500 µL of Blocking/
Storage Buffer. Vortex and sonicate for approximately 20
seconds. Rotate for 30 minutes in the dark at room temperature
to block the coupled microspheres. Centrifuge the microspheres
1 to 2 minutes at ≥ 8,000 × g. Aspirate the supernatant.
6. If you plan to use the coupled microspheres at a later day,
resuspend the microspheres in 500 µL of Wash Buffer. Vortex
and sonicate for approximately 20 seconds.
7. Centrifuge the microspheres 1 minute at ≥ 8,000 × g for 1 to 2
minutes.
8. Aspirate the supernatant. Resuspend the microspheres in 1 mL of
Wash Buffer. Vortex and sonicate for about 20 seconds.
9. Centrifuge the microspheres 1 minute at ≥ 8,000 × g for 1 to 2
minutes.
10. Repeat steps 8 and 9 for a total of two washes with Wash Buffer.
11. Resuspend the coupled and washed microspheres in 500 µL of
Blocking/Storage Buffer.
12. Count the microsphere suspension by hemacytometer.
Calculation: Total microspheres = Count (1 corner of 4 x 4
section) x (1 x 10
4
) x (dilution factor) x (resuspension volume in
mL).
13. Store the preparation at 2
o
C - 8
o
C. Protect it from light.
See note 2.
Technical Notes 1. For best results, use microcentrifuge tubes from USA Scientific,
Inc., catalog number 1415-2500. To order, call 1-800-LAB-TIPS.
2. Coupling can be performed in 100 mM MES, pH 6.0 with
similar results. For some proteins, better solubility and better
coupling may be achieved at a higher coupling pH or in a
- 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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