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Wet transfer is the most common and traditional strategy for western blot transfer due to its efficiency and throughput. We offer three systems for tank transfer that minimize the amount of transfer buffer required.
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Mini Blot Module | XCell II Blot Module | SureLock Tandem Midi Blot Module | |
---|---|---|---|
Capacity | 1 mini gel per blot module; 1-2 blot modules per tank | 1-2 mini gels per blot module; 1 blot module per tank | 1 midi gel per blot module, 1-2 blot modules per tank |
Transfer buffer requirements | 220 mL per blot | 800 mL | 300 mL per blot |
Transfer time | 60 min | 60-120 min | 30 min |
Blotting area | 9 x 9 cm | 9 x 9 cm | 9.2 x 14.4 cm |
Compatible power supply | PowerEase Touch Power Supplies, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) | PowerEase Touch Power Supplies, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) | PowerEase Touch Power Supplies or Owl systems |
Required equipment | Mini Gel Tank | XCell SureLock Mini-Cell or XCell II Mini-Cell | SureLock Tandem Midi Gel Tank: capacity for up to 2 blot modules |
Two Mini Blot Modules fit into the Mini Gel Tank and are designed to make your western transfers simple and easy to perform.
Gel type | Membrane | Voltage | Starting current (mA) | Ending current (mA) | Run time |
---|---|---|---|---|---|
Bolt Bis-Tris Plus 4-12% (MES) | Nitrocellulose | 10 | 160 | 60 | 60 |
PVDF | 20 | 340 | 130 | 60 | |
NuPAGE 4-12% Bis-Tris (MES) | Nitrocellulose | 10 | 160 | 60 | 60 |
PVDF | 20 | 390 | 130 | 60 | |
Novex 4-20% Tris-Glycine (denatured) | Nitrocellulose | 10 | 70 | 50 | 60 |
PVDF | 20 | 160 | 100 | 60 | |
NuPAGE 3-8% Tris Acetate (denatured) | Nitrocellulose | 10 | 150 | 50 | 60 |
PVDF | 20 | 380 | 130 | 60 | |
Novex 10-20% Tricine | Nitrocellulose | 10 | 70 | 60 | 60 |
PVDF | 20 | 180 | 150 | 60 |
* Current readings represent values when running a single gel, and can vary depending upon the power supply being used.
Learn how to perform a western blot transfer using the Mini Blot module in a Mini Gel Tank.
Learn how to separate proteins by using a Invitrogen Bolt Bis-Tris Plus SDS-PAGE precast gel and Mini Gel Tank.
Bolt Western Pack A (Nitrocellulose) | Bolt Western Pack B (PVDF) |
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Includes:
| Includes:
|
The XCell II Blot Module allows you to easily transfer proteins or nucleic acids from mini-gels to membranes. It fits neatly into the XCell SureLock and XCell II Mini-Cells in place of the gel/buffer core assembly. It requires less than 200 mL of transfer buffer for Western, Southern, and northern transfers. Tough platinized titanium and stainless steel electrodes create a uniform electrical field without clamps or hinged gel holders. Maximum blot size is 9 cm x 9 cm.
Gel type | Membrane | Transfer buffer | Transfer conditions | Expected current | Run time |
---|---|---|---|---|---|
Tris-Glycine | Nitrocellulose or PVDF | Tris-Glycine Transfer Buffer with 20% methanol. 1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol. | 25V constant | 100 mA | 1-2 hr |
Tricine | Nitrocellulose or PVDF | Tris-Glycine Transfer Buffer with 20% methanol. 1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol. | 25V constant | Start: 100 mA | 1-2 hr |
Bis-Tris | Nitrocellulose or PVDF | Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples | 30 V constant | Start: 170 mA End: 100mA | 1 hour |
Tris-Acetate | Nitrocellulose or PVDF | Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples | 30 V constant | Start: 200 mA End: 180mA | 60 |
IEF | Nitrocellulose or PVDF | 0.7% acetic acid, pH 3.0 | 10 V constant | Start: 65-85 mA | 60 |
TBE, TBE-Urea and DAN Retardation | Nylon | 45 mM Tris, 45 mM boric acid, 1 mM EDTA | 30 V constant | Start: 360 mA End: 270 mA | 1-2 hr |
The expected current listed in the table below is for transferring one gel. If you are transferring two gels in the blot module, the expected current will double.
For overnight blotting, transfer at a constant voltage of 10-15 V.
Learn how to perform a wet tank transfer using the XCell II Blot Module in the SureLock Tank with this detailed step by step video.
Watch this step by step video of how to run a NuPAGE precast get in the SureLock Tank.
When your western blotting workflow requires higher throughput, Invitrogen midi gels will allow you to load up to 26 samples per gel. You can use the SureLock Tandem Midi Gel Tank to run up to 2 high-performance Invitrogen midi gels and transfer them to membranes in the same tank using the SureLock Tandem Midi Blot Module. Two SureLock Tandem Midi Blot Modules fit into the innovative, dual-purpose SureLock Tandem Midi Gel Tank and are designed for easy, room-temperature gel transfers. Only 300 mL of methanol-containing buffer is required per gel transfer, reducing buffer and hazardous disposal costs.
SureLock Tandem Midi Welcome Pack, Nitrocellulose |
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Includes:
|
When used with high-performance Invitrogen precast midi gels, you can depend on the SureLock Tandem Midi Gel Tank and Midi Blot Module to generate consistent, high-throughput western blots. The Invitrogen midi gels offer reproducible high-performance protein separation, resulting in sharp, well-resolved bands in straight lanes, and the SureLock Tandem Midi Blot Module provides dependable uniform transfer of proteins across the gel.
To demonstrate the SureLock Tandem Midi Blot Module’s uniformity of transfer across a membrane, a NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli lysate ranging from 2-0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard. Electrophoresis was performed and the gel was transferred onto a 0.45 µm PVDF membrane. Results are shown in Figure 1.
Figure 1. Uniform transfer using SureLock Tandem Midi Gel Tank. Total protein normalization of transferred proteins shows uniform transfer across the blot.
A. No-Stain Reagent Labeled PVDF Blot. A NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli Lysate ranging from 2 to 0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard in lanes 1-2 and 19-20. The gel was electrophoresed using MES running buffer in the SureLock Tandem Midi Gel Tank. Proteins from the gel were transferred onto a 0.45 µm PVDF membranes at 25V constant for 30 minutes using the SureLock Tandem Midi Blot Module. After transfer, the proteins on the membrane were labeled following the No-Stain Protein Labeling Reagent protocol for midi-sized membranes. The image was captured using an iBright Imager with the No-Stain Membrane epi setting (455-485 nm excitation and 565-615 nm emission). As the SeeBlue Plus2 Pre-stained Protein Standard is already labeled, its protein bands are not all susceptible to labeling with No-Stain Reagent, and therefore are not all visible in this image.
B. Densitometric Signal Linearity versus Protein Load. The densitometric signal intensity was determined for each lane. Technical replicates (n=4) were averaged and plotted to determine the linear regression for the entire concentration range (R2=0.9994). Error bars represent standard deviation.
Constant Voltage (V) | Time (min) |
---|---|
25 | 30 |
![]() | ![]() | ![]() | |
Mini Blot Module | XCell II Blot Module | SureLock Tandem Midi Blot Module | |
---|---|---|---|
Capacity | 1 mini gel per blot module; 1-2 blot modules per tank | 1-2 mini gels per blot module; 1 blot module per tank | 1 midi gel per blot module, 1-2 blot modules per tank |
Transfer buffer requirements | 220 mL per blot | 800 mL | 300 mL per blot |
Transfer time | 60 min | 60-120 min | 30 min |
Blotting area | 9 x 9 cm | 9 x 9 cm | 9.2 x 14.4 cm |
Compatible power supply | PowerEase Touch Power Supplies, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) | PowerEase Touch Power Supplies, Owl systems, for other systems use with Novex Power Supply Adapters (Cat. No. ZA10001) | PowerEase Touch Power Supplies or Owl systems |
Required equipment | Mini Gel Tank | XCell SureLock Mini-Cell or XCell II Mini-Cell | SureLock Tandem Midi Gel Tank: capacity for up to 2 blot modules |
Two Mini Blot Modules fit into the Mini Gel Tank and are designed to make your western transfers simple and easy to perform.
Gel type | Membrane | Voltage | Starting current (mA) | Ending current (mA) | Run time |
---|---|---|---|---|---|
Bolt Bis-Tris Plus 4-12% (MES) | Nitrocellulose | 10 | 160 | 60 | 60 |
PVDF | 20 | 340 | 130 | 60 | |
NuPAGE 4-12% Bis-Tris (MES) | Nitrocellulose | 10 | 160 | 60 | 60 |
PVDF | 20 | 390 | 130 | 60 | |
Novex 4-20% Tris-Glycine (denatured) | Nitrocellulose | 10 | 70 | 50 | 60 |
PVDF | 20 | 160 | 100 | 60 | |
NuPAGE 3-8% Tris Acetate (denatured) | Nitrocellulose | 10 | 150 | 50 | 60 |
PVDF | 20 | 380 | 130 | 60 | |
Novex 10-20% Tricine | Nitrocellulose | 10 | 70 | 60 | 60 |
PVDF | 20 | 180 | 150 | 60 |
* Current readings represent values when running a single gel, and can vary depending upon the power supply being used.
Learn how to perform a western blot transfer using the Mini Blot module in a Mini Gel Tank.
Learn how to separate proteins by using a Invitrogen Bolt Bis-Tris Plus SDS-PAGE precast gel and Mini Gel Tank.
Bolt Western Pack A (Nitrocellulose) | Bolt Western Pack B (PVDF) |
---|---|
Includes:
| Includes:
|
The XCell II Blot Module allows you to easily transfer proteins or nucleic acids from mini-gels to membranes. It fits neatly into the XCell SureLock and XCell II Mini-Cells in place of the gel/buffer core assembly. It requires less than 200 mL of transfer buffer for Western, Southern, and northern transfers. Tough platinized titanium and stainless steel electrodes create a uniform electrical field without clamps or hinged gel holders. Maximum blot size is 9 cm x 9 cm.
Gel type | Membrane | Transfer buffer | Transfer conditions | Expected current | Run time |
---|---|---|---|---|---|
Tris-Glycine | Nitrocellulose or PVDF | Tris-Glycine Transfer Buffer with 20% methanol. 1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol. | 25V constant | 100 mA | 1-2 hr |
Tricine | Nitrocellulose or PVDF | Tris-Glycine Transfer Buffer with 20% methanol. 1X Transfer Buffer should be pH 8.3 before addition of SDS or methanol. | 25V constant | Start: 100 mA | 1-2 hr |
Bis-Tris | Nitrocellulose or PVDF | Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples | 30 V constant | Start: 170 mA End: 100mA | 1 hour |
Tris-Acetate | Nitrocellulose or PVDF | Bis-Tris transfer buffer with 10% methanol and antioxidant for reduced samples | 30 V constant | Start: 200 mA End: 180mA | 60 |
IEF | Nitrocellulose or PVDF | 0.7% acetic acid, pH 3.0 | 10 V constant | Start: 65-85 mA | 60 |
TBE, TBE-Urea and DAN Retardation | Nylon | 45 mM Tris, 45 mM boric acid, 1 mM EDTA | 30 V constant | Start: 360 mA End: 270 mA | 1-2 hr |
The expected current listed in the table below is for transferring one gel. If you are transferring two gels in the blot module, the expected current will double.
For overnight blotting, transfer at a constant voltage of 10-15 V.
Learn how to perform a wet tank transfer using the XCell II Blot Module in the SureLock Tank with this detailed step by step video.
Watch this step by step video of how to run a NuPAGE precast get in the SureLock Tank.
When your western blotting workflow requires higher throughput, Invitrogen midi gels will allow you to load up to 26 samples per gel. You can use the SureLock Tandem Midi Gel Tank to run up to 2 high-performance Invitrogen midi gels and transfer them to membranes in the same tank using the SureLock Tandem Midi Blot Module. Two SureLock Tandem Midi Blot Modules fit into the innovative, dual-purpose SureLock Tandem Midi Gel Tank and are designed for easy, room-temperature gel transfers. Only 300 mL of methanol-containing buffer is required per gel transfer, reducing buffer and hazardous disposal costs.
SureLock Tandem Midi Welcome Pack, Nitrocellulose |
---|
Includes:
|
When used with high-performance Invitrogen precast midi gels, you can depend on the SureLock Tandem Midi Gel Tank and Midi Blot Module to generate consistent, high-throughput western blots. The Invitrogen midi gels offer reproducible high-performance protein separation, resulting in sharp, well-resolved bands in straight lanes, and the SureLock Tandem Midi Blot Module provides dependable uniform transfer of proteins across the gel.
To demonstrate the SureLock Tandem Midi Blot Module’s uniformity of transfer across a membrane, a NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli lysate ranging from 2-0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard. Electrophoresis was performed and the gel was transferred onto a 0.45 µm PVDF membrane. Results are shown in Figure 1.
Figure 1. Uniform transfer using SureLock Tandem Midi Gel Tank. Total protein normalization of transferred proteins shows uniform transfer across the blot.
A. No-Stain Reagent Labeled PVDF Blot. A NuPAGE 4-12% Bis-Tris Midi gel was loaded with recurring dilutions of E. coli Lysate ranging from 2 to 0.25 µg and SeeBlue Plus2 Pre-stained Protein Standard in lanes 1-2 and 19-20. The gel was electrophoresed using MES running buffer in the SureLock Tandem Midi Gel Tank. Proteins from the gel were transferred onto a 0.45 µm PVDF membranes at 25V constant for 30 minutes using the SureLock Tandem Midi Blot Module. After transfer, the proteins on the membrane were labeled following the No-Stain Protein Labeling Reagent protocol for midi-sized membranes. The image was captured using an iBright Imager with the No-Stain Membrane epi setting (455-485 nm excitation and 565-615 nm emission). As the SeeBlue Plus2 Pre-stained Protein Standard is already labeled, its protein bands are not all susceptible to labeling with No-Stain Reagent, and therefore are not all visible in this image.
B. Densitometric Signal Linearity versus Protein Load. The densitometric signal intensity was determined for each lane. Technical replicates (n=4) were averaged and plotted to determine the linear regression for the entire concentration range (R2=0.9994). Error bars represent standard deviation.
Constant Voltage (V) | Time (min) |
---|---|
25 | 30 |
For Research Use Only. Not for use in diagnostic procedures.