Bio-Rad TGX Stain-Free FastCast Acrylamide Solutions User Manual
TGX™ and TGX Stain-Free™ FastCast™
Acrylamide Kit and Starter Kit Instruction Manual
TGX and TGX Stain-Free FastCast acrylamide kits are solutions for hand casting gels for SDS-PAGE or PAGE use. The unique
formulation enables:
• Ability to cast resolving and stacking gels at the same time and start running your gels in as little as 30 min
• Ability to conduct electrophoretic runs as fast as 20 min (at 300 V) without damage to the handcast gels
• Efficient protein transfers onto blot in as little as 3 min (using the Trans-Blot® Turbo™ blotter)
• 1 year of shelf life at room temperature (22–28°C) for acrylamide solutions and up to 4 weeks at 4ºC for cast gels
• Stain-free visualization of proteins in your gel or blot in less than 5 min with TGX Stain-Free kits
• Ability to monitor your electrophoresis and transfer steps without additional staining using TGX Stain-Free kits
Table 1. Approximate number of gels per kit.
Mini Gels (~9 x 7 cm) Midi Gels (~13 x 9 cm)
1.0 mm
Mini-PROTEAN
Cassettes
Starter Kit 11 gels 19 gels 13 gels 9 gels 6 gels
Kit 55 gels 95 gels 65 gels 45 gels 30 gels
®
0.75 mm
Glass Plates
1.0 mm
Glass Plates
1.5 mm
Glass Plates
When casting different numbers of gels, prepare the solutions according to Table 2 and follow the instructions.
The starter kits enable mixing the stacker A and B and resolver A and B solutions without calculation and casting at least 10
Mini-PROTEAN gels at one time.
Note: After adding APS and TEMED to the resolver solution mixture, casting should be started immediately and combs inserted within 8–9 min. To ensure sufficient
time to insert combs, do not cast more than 10 gels at one time.
Table 2. Casting preparation volumes.
1.0 mm
Mini-PROTEAN
Empty Cassettes
(n = gels)
Stacker Resolver Stacker Resolver Stacker Resolver Stacker Resolver Stacker Resolver
Resolver A – 3.5 ml x n – 2 ml x n – 3 ml x n – 4 ml x n – 6 ml x n
Resolver B – 3.5 ml x n – 2 ml x n – 3 ml x n – 4 ml x n – 6 ml x n
Stacker A 1 ml x n – 1 ml x n – 1 ml x n – 1.5 ml x n – 1.5 ml x n –
Stacker B 1 ml x n – 1 ml x n – 1 ml x n – 1.5 ml x n – 1.5 ml x n –
Total Volume 2 ml x n 7 ml x n 2 ml x n 4 ml x n 2 ml x n 6 ml x n 3 ml x n 8 ml x n 3 ml x n 12 ml x n
0.75 mm
Bio-Rad
Glass Plates
(n = gels)
1.0 mm
Bio-Rad
Glass Plates
(n = gels)
1.0 mm
™
Criterion
Cassettes
1.5 mm
Bio-Rad
Glass Plates
(n = gels)
1.0 mm
Criterion
Empty Cassettes
(n = gels)
TEMED 2 µl x n 3.5 µl x n 2 µl x n 2 µl x n 2 µl x n 3 µl x n 3 µl x n 4 µl x n 3 µl x n 6 µl x n
10% APS 10 µl x n 35 µl x n 10 µl x n 20 µl x n 10 µll x n 30 µl x n 15 µl x n 40 µl x n 15 µl x n 60 µl x n
Instructions
1. Prepare resolving gel acrylamide solution by combining equal volumes of resolver A and B solutions (see Table 2).
Note: To use all the solutions from the starter kits, pour 40 ml resolver B solution directly into the larger amber bottle containing 40 ml resolver A solution.
2. If using the Mini-PROTEAN plastic cassettes, they should be held vertically for gel casting. Use the single-row AnyGel™
casting stand (catalog #165-4131) to stabilize the cassette during casting. For Bio-Rad handcast glass plates, use the
Mini-PROTEAN Tetra cell casting stand (catalog #165-8051 or #165-8052) to stabilize the plates during casting.
3. Add the required volume of TEMED and freshly made 10% APS to the combined resolver solution and mix well. Use an
appropriate pipet to steadily dispense the solution into the cassette. Do not let bubbles form or solution mix with air. Fill the
cassette to 0.5–1 cm below the bottom of the teeth on the comb. Immediately prepare and pour the stacking solution as
directed in the next steps.
Note: Insert a comb and mark the cassette to ensure the resolving solution is poured to the desired distance below the comb.
Note: For the combined resolving gel solutions from the starter kits, add 40 µl of TEMED and 400 µl of freshly made 10% APS.
4. Prepare stacking gel acrylamide solution by combining equal volumes of stacker A and B solutions (see Table 2).
Note: To use all the solutions from the starter kits, pour the 15 ml stacker B solution directly into the larger amber bottle containing 15 ml stacker A solution.
5. Add required volume of TEMED and freshly made 10% APS to the combined stacker solution and mix well. Pipet solution
down the middle of the cassette, filling to the top of the short plates. Apply slowly and steadily to prevent mixing with the
resolving solution. Align and insert the comb in the cassette carefully to prevent air from being trapped under the comb teeth.
Allow the gel to polymerize for 30–45 min before electrophoresis.
Note: For the combined stacking gel solutions from the starter kit, add 30 µl of TEMED and 150 µl of freshly made 10% APS.
6. The cast gels can be covered with a paper towel
Life Science
Group
Sig 121210033918 Rev B US/EG
Bio-Rad
Laboratories, Inc.
Web site ww w.bio-rad.com USA 800 424 6723 Australia 61 2 9914 2800 Austria 01 877 89 01 Belgium 09 385 55 11 Brazil 55 11 5044 56 99
Canada 905 364 3435 China 86 21 6169 8500 Czech Republic 420 241 430 532 Den mark 44 52 10 00 Finland 09 80 4 22 00
France 01 47 95 69 65 Germany 089 31 884 0 Greece 30 210 953 2 220 Hong Kon g 852 2789 330 0 Hungary 36 1 459 6100 India 91 124 4029300
Israel 03 963 605 0 Italy 39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460 Mexico 52 555 4 88 7670 The Netherlands 0318 540666
New Zealand 64 9 415 2280 No rway 23 38 41 30 Poland 48 22 3 31 99 99 Portugal 351 21 472 7700 Russia 7 495 721 14 04
Singapore 65 6415 3188 South Africa 27 861 246 723 Spain 34 91 590 52 00 Swed en 08 555 12700 Switzerland 026 674 55 05
Taiwan 886 2 2578 7189 Thailand 800 88 22 88 United Kingdom 020 8328 2 000
wetted with deionized water and stored in a zipper
sealed plastic bag at 4°C for up to 4 weeks.
7. Follow the guideline below for gel electrophoresis,
stain-free imaging, and western blot transfer. Refer to
the Mini-PROTEAN Precast Gels Instruction Manual
and Application Guide (#1658100) for details on
running, handling, and other applications of TGX and
TGX Stain-Free gels.
Use Laemmli sample buffer (with or without reducing
agents such as BME) to prepare samples for loading
and 1x Tris/glycine/SDS running buffer to run the TGX
gels.
Table 3. Running conditions for handcast TGX or TGX Stain-Free
SDS-PAGE gels in the Mini-PROTEAN Tetra cell.
100 V
Low voltage
Run time
Lower chamber buffer
volume (for 2 gels)
Lower chamber buffer
volume (for 4 gels)
Note:
• When running only 1–2 gels in the Mini-PROTEAN Tetra cell, do not
leave the companion module in the tank
• Do not run different gel types (chemistry) or percentages in the same
tank at the same time
Table 4. Stain-Free gel activation using a Bio-Rad Stain-Free enabled
imager.
Activation time
* For gels not used in blotting.
Note:
• Must activate stain-free compound in gel for stain-free blot imaging
• Stain-free activation protocols can be accessed in Bio-Rad’s
Image Lab™ software
Table 5. Transfer conditions for blotting in the Trans-Blot Turbo
system.
Protocol
Name
Standard SD Any 30 Up to 1.0 A; 25 V, constant
1.5 mm gel Any 10
High MW >150 10
Low MW <30 5
Mixed MW* 5–150 7
1 Mini TGX* 5–150 3 N/A
* Also accessed via Turbo navigation button.
Precision Plus Protein standards are sold under license from Life Technologies Corporation, Carlsbad,
CA, for use only by the buyer of the product. The buyer is not authorized to sell or resell this product or its
components.
MW, kD Time, min 2 Mini Gels or
85–95 min 30–40 min 20–25 min
550 ml 550 ml 800 ml
800 ml 800 ml 800 ml
Gels used in
Blotting
1 min 2.5 min 5 min
200 V
Standard
Good*
Sensitivity
1 Midi Gel
2.5 A constant
up to 25 V
300 V
Rapid
Best*
Sensitivity
1 Mini Gel
1.3 A constant
up to 25 V
2.5 A constant
up to 25 V
Ordering Information
Catalog # Description
161-0170 TGX™ FastCast™ Acrylamide Starter Kit, 7.5%
161-0171 TGX FastCast Acrylamide Kit, 7.5%
161-0172 TGX FastCast Acrylamide Starter Kit, 10%
161-0173 TGX FastCast Acrylamide Kit, 10%
161-0174 TGX FastCast Acrylamide Starter Kit, 12%
161-0175 TGX FastCast Acrylamide Kit, 12%
161-0180 TGX Stain-Free™ FastCast™ Acrylamide Starter Kit, 7.5%
161-0181 TGX Stain-Free FastCast Acrylamide Kit, 7.5%
161-0182 TGX Stain-Free FastCast Acrylamide Starter Kit, 10%
161-0183 TGX Stain-Free FastCast Acrylamide Kit, 10%
161-0184 TGX Stain-Free FastCast Acrylamide Starter Kit, 12%
161-0185 TGX Stain-Free FastCast Acrylamide Kit, 12%
161-0700 Ammonium Persulfate (APS), 10 g
161-0800 TEMED, 5 ml
All cell systems are 4-gel with 2 casting stands, 4 casting frames,
10-well combs, 5 short plates, and 5 spacer plates.
165-8000 Mini-PROTEAN® Tetra Cell and System,
0.75 mm gel thickness
165-8001 Mini-PROTEAN Tetra Cell and System,
1.0 mm gel thickness
165-8006 Mini-PROTEAN Tetra Cell and System,
1.5 mm gel thickness
Standards Value Pack (5 Packs)
161-0393 Precision Plus Protein™ All Blue Standards; 10–250 kD
161-0394 Precision Plus Protein Dual Color Standards; 10–250 kD
161-0395 Precision Plus Protein™ Kaleidoscope™ Standards; 10–250 kD
161-0397 Precision Plus Protein Dual Xtra Standards; 2–250 kD
161-0396 Precision Plus Protein Unstained Standards; 10–250 kD
161-0737 2x Laemmli Sample Buffer, 30 ml
161-0747 4x Laemmli Sample Buffer, 10 ml
161-0772 10x Tris/Glycine/SDS Buffer, 5 L
Mini-PROTEAN Empty Cassettes and Combs, pkg of 50
456-0001 Mini-PROTEAN Empty Cassettes, IPG well marks
456-0003 Mini-PROTEAN Empty Cassettes, 10-well marks
456-0005 Mini-PROTEAN Empty Cassettes, 12-well marks
456-0006 Mini-PROTEAN Empty Cassettes, 15-well marks
456-0011 Mini-PROTEAN Combs, IPG well combs
456-0013 Mini-PROTEAN Combs, 10-well combs
456-0015 Mini-PROTEAN Combs, 12-well combs
456-0016 Mini-PROTEAN Combs, 15-well combs
Criterion Empty Cassettes, pkg of 10 cassettes with combs
345-9901 Criterion Empty Cassettes, 12+2 well
345-9902 Criterion Empty Cassettes, 18-well
345-9903 Criterion Empty Cassettes, 26-well
345-9904 Criterion Empty Cassettes, Prep+2
345-9906 Criterion Empty Cassettes, IPG+1
* Please see the accessories tab on www.bio-rad.com/tetra for individual
handcast products.
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