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PN 89-00002-00-084 Rev. B 59
xMAP Technology LumAvidin Binding Protocol
LumAvidin Binding Protocol
Introduction Use these protocols as a general starting point for developing assays.
Optimize all assays for your reagents in your specific application.
Updates or additions to these protocols are posted on the Luminex
website at
http://luminexcorp.custhelp.com. At the main page, select
a subject or perform a search for the desired information
You obtain the best results by starting with these guidelines and
modifying them for your specific needs.
Equipment •Vortex
• Micropipetters (1 µL - 1000 µL)
• Microcentrifuge
•Timer
•Rotator
Materials • xMAP LumAvidin-modified xMAP microspheres—LIMIT
EXPOSURE TO LIGHT!
• Biotin-conjugated molecule
• Microcentrifuge tubes: 1.5 mL, polypropylene (see technical
note 3)
• REACTION BUFFER:
Phosphate Buffered Saline (pH 7.3 ± 1%), Bovine Serum
Albumin (BSA)
• BLOCKING/STORAGE BUFFER:
Phosphate Buffered Saline (pH 7.3 ± 0.1), Tween 20
(0.02% v/v), Bovine Serum Albumin (1 mg/mL), Sodium Azide
(0.05% w/v)
Preparation Dilute the Biotin-conjugated molecule in REACTION BUFFER to a
concentration of 4 - 4000 nM (See Technical note 1).
Procedure 1. Centrifuge the Avidin-modified xMAP microsphere stock for 1
minute at ≥ 8,000 × g.
2. Disperse the pellet with sonication, and vortex the container for
20 seconds.
3. Dispense 1.0 × 10
5
Avidin-modified xMAP microspheres into a
1.5 mL USA Scientific microcentrifuge tube.
4. Centrifuge the xMAP microspheres for 1 minute at ≥ 8,000 × g.
™
Note: We recommend titration
in the 4 to 4000 nM range to
determine the optimal amount of
biotin-conjugated molecule per
specific binding reaction.
- 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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