Bio-Rad TGX Stain-Free™ FastCast™ Acrylamide Solutions - 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
TGX ™ and TGX Stain-Free ™ FastCast ™
Acrylamide Kit and Starter Kit Instruction Manual
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
0.75 mm
Glass Plates
1.0 mm
Glass Plates
1.5 mm
Glass Plates
1.0 mm
Criterion ™
Cassettes
Starter Kit 11 gels 19 gels 13 gels 9 gels 6 gels
Kit 55 gels 95 gels 65 gels 45 gels 30 gels
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)
0.75 mm
Bio-Rad
Glass Plates
(n = gels)
1.0 mm
Bio-Rad
Glass Plates
(n = gels)
1.5 mm
Bio-Rad
Glass Plates
(n = gels)
1.0 mm
Criterion
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
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.