BIO BENCH AP-GPG-610 Precast PAGE Gel Bis Tris and Tris Glycin User Guide

BIO BENCH AP-GPG-610 Precast PAGE Gel Bis Tris and Tris Glycin

Product Information

Introduction

The AccuPAGETM Tris-Gly gel is made of an optimized formula from conventional Tris-Glycine gel. It has good stability and a long shelf life of up to 12 months. If your lab has been using Tris-Glycine gel, then you can shift to AccuPAGETM Tris-Gly precast gel smoothly.

Key Features and Advantages

1. Wide range of protein size adaptability from 3.5kDa to 300kDa.

Specifications of the AccuPAGETM Precast Gel

Gel Type| Concentration| Gel Dimension| Cassette Dimension| Gel Thickness| Stacking Gel| Well Amount and Max Load| Storage and Shelf Life| Tank Compatibility| Running Time| Application| Running Buffer| Quantity
---|---|---|---|---|---|---|---|---|---|---|---|---
Tris-Glycine Gel| 6%, 8%, 10%, 12%, 15%, 4-15%, 4-20%| 1mm| 10 wells 50L, or 15 wells 30L| 10| Tris-Glycine| Native PAGE, denaturing PAGE. Note: Gel does NOT contain
detergent (like SDS).| Room temperature or controlled temperature.| Tank Compatibility| Running Time| Application| Tris-Glycine| 2 pcs test pack, 10 pcs, 30 pcs and 50 pcs
Bis-Tris Gel| 8%, 10%, 12%, 4-12%| 1mm| 10 wells 50L, or 15 wells 30L| 10| MOPS or MES| Native PAGE, denaturing PAGE. Note: Gel does NOT contain
detergent (like SDS).| Room temperature or controlled temperature.| Tank Compatibility| Running Time| Application| MOPS or MES| 2 pcs test pack, 10 pcs, 30 pcs and 50 pcs

Protein Separation using AccuPAGETM Precast Gels

The chart below shows the relative migration of proteins in denatured condition. Please check this chart before choosing the gel.

AccuPAGETM Precast Gel List and Accessories

Product Usage Instructions

Running Buffer Preparation

To prepare the running buffer, follow these steps:

1. Choose the appropriate running buffer based on the application.
2. Prepare the running buffer according to the manufacturer’s instructions.

Sample Preparation

To prepare the sample for protein separation, follow these steps:

1. Prepare the protein sample according to your experimental requirements.
2. Add any necessary loading buffer or reducing agent to the protein sample.
3. Mix the sample gently to ensure proper mixing of the components.

Instructions for Using AccuPAGETM Precast Gel

To use the AccuPAGETM Precast Gel, follow these steps:

1. Open the gel cassette carefully and remove any protective coverings.
2. Place the gel cassette in the appropriate gel tank or electrophoresis system.
3. Add the running buffer to the gel tank according to the manufacturer’s instructions.
4. Load the prepared protein sample into the wells of the gel using a pipette or loading device.
5. Close the gel cassette securely to prevent any leakage during the electrophoresis process.
6. Connect the gel tank to a power supply and set the appropriate voltage and running time.
7. Start the electrophoresis process and monitor the progress as needed.
8. After the electrophoresis is complete, carefully remove the gel cassette from the tank.

Troubleshooting Guide

Refer to the troubleshooting guide in the user manual for assistance with any issues you may encounter during product usage.

Notice
Read and follow all safety precautions and guidelines provided in the user manual before using the product.

Contact Information
For any inquiries or support, please contact our customer service department at [email protected]

Standard Warranty
This product is covered by our standard warranty. Please refer to the warranty terms and conditions in the user manual for more information.

Introduction

Bio Bench AccuPAGE™ protein electrophoresis gels are precast polyacrylamide gels that provide large range of molecular weight separation ability with good resolution and sample integrity. AccuPAGE™ provides 2 types of gels, including conventional Tris-Glycine gel and the Bis-Tris gel. The formula of these 2 types of gel are both optimized by Bio Bench, in order to provide better performance as well as good stability during long term storage. AccuPAGE™ is compatible with nearly all of tanks from major biotech providers. Take out from the fridge and open the package, then you can operate it under the routine lab instructions.

AccuPAGE™ Tris-Gly gel is made of optimized formula from conventional Tris- Glycine gel. It has good stability and long shelf life for up to 12 months. If you lab has been using Tris-Glycine gel, then you can shift to AccuPAGE™ Tris- Gly precast gel smooth.
AccuPAGE™ Bis-Tris gel delivers consistent gel performance with a neutral pH environment (pH≈7.0) that minimizes any protein modifications. Bis-Tris can improve the stability of the gel structure during electrophoresis, which will avoid the gel become trapezoid shape and lead to sharp and straight band after staining. Your lab device does not need to be changed to run AccuPAGE™ Bis- Tris gel – only to remember that Bis-Tris gel needs to be run in either MOPS or MES buffer.

Key features and advantages

1. Wide range of protein size adaptability from 3.5kDa to 300kDa. AccuPAGE™ Precast Gels are available in different acrylamide concentrations, including 6%, 8%,10%,12% and 4-12%, 4-15% and 4-20%. Customized service for special concentration is also available.
2. Good compatibility with electrophoresis tanks from almost all brands, including but not limited to: Bio-Rad, Life Technology (Thermo), Hoefer, Sigma Aldrich Dual Run, Thermo XCell I, II, & Surelock™ mini-cell, LONZA PAGEr® Minigel Chamber and Tanon, etc.
3. Performance tested in-house by Bio Bench’s CRO services. We use it in our lab!
4. Stability and repeatability of PAGE result ensured by automated gel casting platform.
5. Coated plastic cassettes avoid protein absorption and ensures the clear and sharp protein band.
6. Rapid electrophoresis within 25min by Ultra Fast Running Buffer.
7. Compatible with all types of protein marker, including conventional marker and pre-stained marker.
8. The gel can be observed by Coomassie brilliant blue staining, silver staining, fluorescent staining or by UV light.
9. Gel cassettes can be easily opened by the common Gel Opener.
10. AccuPAGE™ Precast Gels is free of any detergent (like SDS). User can run either native PAGE or denaturing PAGE.

Specifications of the AccuPAGE™ Precast Gel

4-20%

| ****

8%, 10%, 12%, 4-12%

Gel Dimension| Mini gel, 84mm(W) × 74mm(H) × 1mm(D)
Cassette Dimension| 100mm(W) × 89mm(H) × 4.8mm(D)
Gel Thickness| 1mm
Stacking Gel| Concentration 4%, 15mm(H)
Well amount and max load| ****

10 wells 50μL, or 15 wells 30μL.

Storage and Shelf Life Shipping| 4 – 8°C, up to 12 months.

DO NOT FREEZE.

| 4 – 8°C, up to 12 months.

DO NOT FREEZE.

Tank Compatibility| Room temperature or controlled temperature.

Tank Compatibility

| ****

AccuPAGE™ is designed to be compatible with all types of MINI-PAGE tank which allows 10cm width gel cassette. Including but not limited to the following models:

1. Bio-Rad Mini-PROTEAN (II/3 /Tetra System) *

2. Hoefer™ Mighty Small (SE250 / SE260 / SE280)

3. Sigma Aldrich Dual Run**

4. Thermofisher/Invitrogen XCell (I, II, & Surelock™ mini-cell)**

5. LONZA PAGEr® Minigel Chamber**

6. Tanon

Running Time| Typically 40 – 60 min when using common buffer.

Minimum 20min when using Ultra Fast Running Buffer.

Application| Native PAGE, denaturing PAGE.

Note: Gel does NOT contain detergent (like SDS).

Running Buffer| Tris-Glycine| MOPS or MES
Quantity| 2 pcs test pack, 10 pcs, 30 pcs and 50 pcs.

• Adaption will be needed, please read Instruction of operation – 4.1.
• Accessory will be needed. please read Instruction of operation – 4.2 and 4.3.

Storage and Stability
AccuPAGE™ Precast Gels have extended shelf-life of up to 12 months from the date of production, when stored at 4–8 °C.
AccuPAGE™ Precast Gels shall be laid flat during the storage. “UP” arrow shows the direction of the box during storage.

Protein Separation using AccuPAGE™ Precast Gels
The chart under is showing the relative migration of the protein in denatured condition. Please check this chart before choosing the gel.

• AccuPAGE™ Bis-Tris Precast Gel, denatured condition
• AccuPAGE™ Tris-Glycine Precast Gel, denatured condition

AccuPAGE™ Precast Gel List and Accessories

Gel type

| Running Buffer| Gel Thickness| Gel Concentration| Max Separation| Recommended Separation| Well Amount| Max Sample Load| ****

Cat. No.

---|---|---|---|---|---|---|---|---

Tris- Glycine

| ****

Tris-Glycine

| ****

1mm

| ****

6%

| ****

40-300kDa

| ****

60-200kDa

| 10| 50μL| AP-GPG-610
15| 30μL| AP-GPG-615

8%

| ****

25-300kDa

| ****

40-200kDa

| 10| 50μL| AP-GPG-810
15| 30μL| AP-GPG-815

10%

| ****

11-230kDa

| ****

20-160kDa

| 10| 50μL| AP-GPG-1010
15| 30μL| AP-GPG-1015
12%| ****

6.5-200kDa

| ****

15-85kDa

| 10| 50μL| AP-GPG-1210
15| 30μL| AP-GPG-1215
15%| 5-170kDa| 10-50kDa| 10| 50μL| AP-GPG-1510
15| 30μL| AP-GPG-1515
4% – 15%| 5-270kDa| 20-200kDa| 10| 50μL| AP-GPG-41510
15| 30μL| AP-GPG-41515
4% – 20%| 3.5-300kDa| 5-200kDa| 10| 50μL| AP-GPG-42010
15| 30μL| AP-GPG-42015

Tris- Bis

| ****

MOPS

or MES

| ****

1mm

| 8%| 8-270kDa| 25-245kDa| 10| 50μL| AP-GPB-810
15| 30μL| AP-GPB-815
10%| 8-270kDa| 11-180kDa| 10| 50μL| AP-GPB-1010
15| 30μL| AP-GPB-1015
12%| 3.5-245kDa| 11-135kDa| 10| 50μL| AP-GPB-1210
15| 30μL| AP-GPB-1215
4% – 12%| 3.5-270kDa| 11-245kDa| 10| 50μL| AP-GPB-41210
15| 30μL| AP-GPB-41215
Product Name| Description| Cat. No.
---|---|---
Tank Adapter Plate – Type 1| Tank Adapter designed for the following models:

1. Sigma Aldrich Dual Run *

2. Thermofisher/Invitrogen XCell (I, II, & Surelock™ mini-cell) 3.LONZA PAGEr® Minigel Chamber

| ****

AP-AC-01

Tank Adapter Plate – Type 2| Tank Adapter designed for the following model:

1. Life Technologies Mini Gel Tank

| ****

AP-AC-02

Ultra Fast Running Buffer for Tris-Gly Gel| Ultra Fast Running Buffer designed for Tris-Gly gel, running

time as little as 20min. 1 pack for 500mL running buffer.

| ****

AP-B-P500

Instructions for Using AccuPAGE™ Precast Gel

Running Buffer Preparation
10× Stock Solution

MOPS, SDS Free

10× stock solution, 1L

| MES, SDS Free

10× stock solution, 1L

| Tris-Glycine, SDS Free 10× stock solution, 1L
---|---|---
50 mM MOPS, 50 mM Tris

Base, 1 mM EDTA, pH 7.7

| 50 mM MES, 50 mM Tris

Base, 1 mM EDTA, pH 7.3

| 1.92 M Glycine, 250mM Tris

Base, pH 8.5

1. Dissolve the following reagents

in 900 mL ultrapure water.

2. Mix well and adjust the volume to 1L with ultrapure water.

3. Do NOT use acid or base to adjust the pH.

| 1. Dissolve the following reagents

in 900 mL ultrapure water.

**** 2. Mix well and adjust the volume to 1L with ultrapure water.

3. Do NOT use acid or base to adjust the pH.

| 1. Dissolve the following reagents

in 900 mL ultrapure water.

2. Mix well and adjust the volume to 1L with ultrapure water.

3. Do NOT use acid or base to adjust the pH.

Add 10g SDS to the stock

solution for denaturing condition.

| Add 10g SDS to the stock

solution for denaturing condition.

| Add 10g SDS to the stock

solution for denaturing condition.

Buffer is stable for 6 months when stored at 4°C.| Buffer is stable for 6 months when stored at 4°C.| Buffer is stable for 6 months when stored at 4°C.

How to Make 1× Working Solution

MOPS

1× working solution, 1L

| MOPS

1× working solution, 1L

| Tris- Glycine

1× working solution, 1L

---|---|---
Mix 100mL of the 10× stock solution with 900mL of ultrapure water.| Mix 100mL of the 10× stock solution with 900mL of ultrapure water.| Mix 100mL of the 10× stock solution with 900mL of ultrapure water.
Buffer is stable for 1 week when stored at 4°C.| Buffer is stable for 1 week when stored at 4°C.| Buffer is stable for 1 week when stored at 4°C.

Running Buffer Selection Guide

Gel

Buffer

| Bis-Tris Denatured| ****

Tris-Gly Denatured

| ****

Bis-Tris Native

| ****

Tris-Gly Native

---|---|---|---|---
Tris-Glycine| ****

DO NOT USE.

| YES

Add Detergent

| ****

DO NOT USE.

| YES

AVOID Detergent !

MOPS| YES

Add Detergent

| DO NOT USE.| YES

AVOID Detergent !

| DO NOT USE.
MES| YES

Add Detergent

| ****

DO NOT USE.

| YES

AVOID Detergent !

| ****

DO NOT USE.

Ultra Fast Running Buffer

Cat: AP-B-P500

| DO NOT USE.| YES

Add Detergent

| DO NOT USE.| DO NOT USE.

Sample Preparation
Sample and Sample Loading Buffer Volume

| ****

**Max Sample Volume***

| ****

2×Sample Loading Buffer

| ****

5×Sample Loading Buffer

| ****

10×Sample Loading Buffer

| ****

β-mercaptoetha- nol, 1M stock**

| ****

10 well precast gel final volume

| ****

15 well precast gel final volume

---|---|---|---|---|---|---|---
Denatured Sample| ****

12μL

| ****

15μL

| ****

\

| ****

\

| ****

3μL

| ****

\

| ****

30μL

Denatured Sample| ****

21μL

| ****

\

| ****

6μL

| ****

\

| ****

3μL

| ****

\

| ****

30μL

Denatured Sample| ****

24μL

| ****

\

| ****

\

| ****

3μL

| ****

3μL

| ****

\

| ****

30μL

Denatured Sample| ****

20μL

| ****

25μL

| ****

\

| ****

\

| ****

5μL

| ****

50μL

| ****

\

Denatured Sample| ****

35μL

| ****

\

| ****

10μL

| ****

\

| ****

5μL

| ****

50μL

| ****

\

Denatured Sample| ****

40μL

| ****

\

| ****

\

| ****

5μL

| ****

5μL

| ****

50μL

| ****

\

Native Sample| ****

15μL

| ****

15μL

| ****

\

| ****

\

| ****

\

| ****

\

| ****

30μL

Native Sample| ****

24μL

| | ****

6μL

| ****

\

| ****

\

| ****

\

| ****

30μL

Native Sample| ****

27μL

| | | 3μL| ****

\

| ****

\

| ****

30μL

Native Sample| ****

25μL

| 25μL| | | | 50μL| ****

\

Native Sample

| 40μL| | 10μL| | | 50μL| \
Native Sample| 45μL| | | 5μL| | 50μL| \

1. The sample volume shall be adjusted according to your experiment. If necessary, user can use different volume of the samples and add purified water till the indicated volume.
2. Either dithiothreitol (DTT) or β-mercaptoethanol (BME) can be used as a reducing/denaturing agent (DTT to a final concentration of 100 mM or add BME to a final concentration of 100 mM).

Instruction of operation

1. Open
   Open the package from the indicated mark.

2. Peel off
   Peel off the sealing tape from the bottom of the gel cassette.

3. Remove
   Gently remove the comb from the gel cassette.

4. Gel installation
   Install the gel cassette in the tank according to manufacturer’s instruction.
   NOTE: some brands or models of the tank need accessories for installation!Read below.

Gel installation – Continue, if using these brands of tank:

• Bio-Rad Mini-PROTEAN® II and 3
• Bio-Rad Mini-PROTEAN® Tetra System
  Take out the green silicon gasket. Reverse to its back and reinstall it back into the frame. Then the AccuPAGE™ Precast Gel can be well attached after installation.

Gel installation – Continue, if using these brands of tank:

• Sigma-Aldrich® Dual Run and Blot System
• Thermo XCell I, II, & Surelock™ mini-cell
• LONZA PAGEr® Minigel Chamber

These tanks are designed for thicker gel cassettes. The AccuPAGE™ Tank Adapter Plate – type 1 (Cat: AP-AC-01) shall be used together with the AccuPAGE™ Precast Gel to fill the gap. Use one piece of Adapter Plate per precast gel.
After electrophoresis, please wash the adaptor plate.

Gel installation – Continue, if using these brands of tank:

Life Technologies Mini Gel Tank

The AccuPAGE™ Tank Adapter Plate – Type 2 (Cat: AP-AC-02) shall be used together with the AccuPAGE™ Precast Gel for successful installation. Use one piece of Adapter Plate per precast gel.

Remove the black rubber from Life technologies Mini Gel tank.

Take out the adaptor plate. User will find both sides of the adaptor plate have black color sealers. One side is longer and it is defined “the long side”; the other side is defined “the short side”.

The long side faces the operator, then place the adaptor plate into the chamber of the tank.

Then install the precast gel behind the adaptor, on the short side. Note that, the sample wells shall face the operator. (Comb is shown in the image to indicate the direction. Please remove the comb before your operation).

Install the cassette clamp, then the gel is successfully installed in the tank.
The precast gel can be installed on both chambers of the Mini Gel tank. After electrophoresis, please wash the adaptor plate.

Fill
Firstly, fill the electrode assembly with 1X running buffer to verify for the proper sealing. Then filling the anode (outer) chamber to the recommended level.

Load
To load the samples into the wells, insert tip vertically for better sample loading. DO NOT EXCEED WELL CAPACITY: 50μL for 10-well gels; and 30 μL for 15-well gels.

Run the gel
Once the samples are loaded and the buffer is filled in the tank, place the cover over the tank and connect the cable to the power supply.

1. Run the gel – continue
   The chart hereunder shows the voltage options for running the AccuPAGE™ Precast Gel. The voltages show as ranges, you may first test according to your lab protocol then adjust the voltage according to the PAGE result. During the running process, the voltage shall not be changed.
   It is ideal to run PAGE at the temperature range of 10°C – 20°C. However, during the run, the electric current will increase and more heat will be generated. If the temperature of the running buffer goes above 30°C, please put the tank into ice bath to keep cool.

| Running Buffer| Typical Voltage| Typical Run time| Optional Voltage| Optional Run time
---|---|---|---|---|---

Bis-Tris Gel

General condition

|

MOPS

|

150V-200V

|

25-50min

|

\

|

\

Bis-Tris Gel

To obtain more clear and sharp bands

|

MOPS

|

\

|

\

|

100V-150V

|

45-60min

Bis-Tris Gel

General condition

|

MES

|

150V-180V

|

25-45min

|

\

|

\

Bis-Tris Gel

To obtain more clear and sharp bands

|

MES

|

\

|

\

|

100V-150V

|

40-60min

Tris-Glycine Gel

General Condition

|

Tris-Gly

|

160-180V

|

50-60min

|

\

|

\

Tris-Glycine Gel

To obtain more clear and sharp bands

|

Tris-Gly

|

\

|

\

|

120V-150V

|

60-90min

Tris-Glycine Gel

General Condition

| Ultra Fast Running Buffer

AP-B-P500

|

250V

|

18-25min

|

\

|

\

Stop
Stop the power supply when the blue staining line is 2mm from the edge. You may stop the power earlier or later in order to optimize your experiment.

Open
Open the cassette to take out the gel.
Once the electrophoresis is finished, user can take out the gel cassette from the electrode assembly. Use the Gel Cassette Opener to open the gel cassette, from the corners as well as top and bottom. OPERATE WITH CAUTION!
After the cassette cover is opened, use the Gel Opener to cut softly the left and right side of the gel border to remove the gel.
If necessary, put the gel cassette in water flow or water bath to help you open the cassette and remove the gel.

Staining
AccuPAGE™ Precast Gel is compatible with common staining protocols. User can use either lab-made staining or commercial staining. Please operate the staining under your lab protocol or the commercial instruction.

Western Blot.
AccuPAGE™ Precast Gel is designed to adapt with popular WB protocols, including both wet transfer and semi-dry transfer. In order to achieve good test result, please follow the blotting equipment manufacturer’s instruction.

Notes to help you to achieve good result:

1. Always use pre-stained marker during transfer. It will let you see the transfer result directly.
2. Both PVDF and NC membranes are compatible with AccuPAGE™ Precast Gel.
3. Do NOT reuse the transfer buffer while following wet transfer protocol.
4. Avoid short circuit of the power supply. Make sure that: membrane is smaller than the gel, filter paper is smaller than the membrane, foam (during wet transfer) is smaller than the filter paper.

Troubleshooting Guide

Notice
All of the information and advices provided in the user guide are based on the best of our knowledge and ability, but without obligation or liability. Our information and advice do not exempt our customers own responsibility for checking the suitability of the products for intended purpose.
The information in the user guide is subjected to change from time to time, without prior notice and should not be entitled as a commitment by manufacturing or selling entity, or an affiliate. We deny any responsibility for any errors that may appear in this document.

Contact information
Please contact the sales agency from where you have purchased the product.
Or you may send email to info@bio-bench.com for any question.

Standard Warranty
12 months warranty from the date of production.
Manual Version
V1.3, 01-12-2022

BIO BENCH – Boost research.
Web: www.bio-bench.com
Email: info@bio-bench.com

References

• Benchmark | Advanced Electronics Engineering & Manufacturing
• BIO BENCH - YOUR RELIABLE CRO PARTNER! - Bio Bench Home Page

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