ProtoGel 40% (EC-891) SDS-PAGE Protocol
ProtoGel 0.4, also known as ProtoGel 40% (EC-891), is a stabilized 40% acrylamide/bis-acrylamide solution with a 37.5:1 acrylamide-to-bisacrylamide ratio, developed for reproducible polyacrylamide gel preparation and protein electrophoresis.
Designed for Laemmli SDS-PAGE workflows, ProtoGel 40% simplifies gel casting by providing a ready-to-use monomer solution for preparing resolving and stacking gels at the required acrylamide percentage.
- Catalog number : EC-891
- Product : ProtoGel 40% / ProtoGel 0.4
- Manufacturer : National Diagnostics
- Format : Stabilized acrylamide/bis-acrylamide solution
- Ratio : 37.5:1 acrylamide : bis-acrylamide
- Applications : SDS-PAGE, protein electrophoresis and protein separation
- Available sizes : 450 mL and 1 L
- CTA : Check ProtoGel 0.4 Availability / Request a Quote
What Is ProtoGel 40% ?
ProtoGel 40% is a concentrated acrylamide/bis-acrylamide solution used to prepare polyacrylamide gels for protein electrophoresis.
Instead of weighing and combining acrylamide and bis-acrylamide monomers separately, researchers can dilute ProtoGel to the desired final gel percentage and combine it with the appropriate resolving or stacking buffer.
Its 37.5:1 acrylamide-to-bisacrylamide ratio is suitable for conventional Laemmli SDS-PAGE protein separation.
Key Benefits of ProtoGel 40%
- Ready-to-use stabilized acrylamide/bis-acrylamide solution.
- 40% concentrated formulation for flexible gel percentages.
- 37.5:1 acrylamide-to-bisacrylamide ratio.
- Optimized for Laemmli SDS-PAGE.
- Helps reduce variability associated with manual monomer preparation.
- Designed to produce clear, reproducible polyacrylamide gels.
- Suitable for protein separation before downstream workflows such as Western blotting.
- Available as catalog number EC-891.
ProtoGel 40% SDS-PAGE Protocol
The amount of ProtoGel 40% required depends on the final acrylamide percentage and total volume of the resolving gel.
ProtoGel Calculation Formula :
=> ProtoGel volume (mL) = Desired gel % × Total gel volume (mL) ÷ 40
For example, to prepare 10 mL of a 10% resolving gel :
=> 10 × 10 ÷ 40 = 2.5 mL ProtoGel 40%
This makes it easy to calculate ProtoGel requirements for different gel concentrations and casting volumes.
ProtoGel 40% Resolving Gel Recipe - 10 mL
| Final Gel | ProtoGel 40% | 4X Resolving Buffer | DI Water | 10% APS | TEMED |
|---|---|---|---|---|---|
| 6% | 1.50 mL | 2.50 mL | 5.89 mL | 0.10 mL | 0.01 mL |
| 8% | 2.00 mL | 2.50 mL | 5.39 mL | 0.10 mL | 0.01 mL |
| 10% | 2.50 mL | 2.50 mL | 4.89 mL | 0.10 mL | 0.01 mL |
| 12% | 3.00 mL | 2.50 mL | 4.39 mL | 0.10 mL | 0.01 mL |
| 15% | 3.75 mL | 2.50 mL | 3.64 mL | 0.10 mL | 0.01 mL |
Resolving Gel Procedure
- Combine ProtoGel 40%, resolving buffer and deionized water according to the required gel percentage.
- For optimal polymerization, degas the gel solution under vacuum for approximately 10 minutes.
- Add freshly prepared 10% ammonium persulfate.
- Mix gently.
- Add TEMED and mix gently again.
- Immediately pour the resolving gel into the casting cassette.
- Allow approximately 10–20 minutes for polymerization.
- After the resolving gel has polymerized, prepare the stacking gel.
APS and TEMED should be added immediately before casting because they initiate acrylamide polymerization.
4% ProtoGel Stacking Gel - 10 mL
For a standard 10 mL 4% stacking gel:
| Component | Volume |
| ProtoGel 40% | 1.00 mL |
| ProtoGel Stacking Buffer | 2.50 mL |
| Deionized water | 6.44 mL |
| 10% APS | 0.05 mL |
| TEMED | 0.01 mL |
After adding APS and TEMED, pour the stacking gel and insert the appropriate comb. Polymerization generally begins within approximately 20 minutes.
Which Running Buffer Should Be Used with ProtoGel?
For standard Laemmli SDS-PAGE, a 1X Tris-Glycine-SDS running buffer is suitable for most protein electrophoresis applications.
For proteins below approximately 20 kDa, a Tris-Tricine-SDS system can provide improved separation of smaller proteins.
The appropriate electrophoresis conditions should always be selected according to the target protein, gel percentage, equipment and validated laboratory protocol.
What Gel Percentage Should I Use?
The optimum acrylamide percentage depends primarily on the molecular weight range of the proteins being separated.
Lower-percentage gels have larger pores and are generally useful for larger proteins, while higher-percentage gels have smaller pores and provide greater separation of smaller proteins.
ProtoGel 40% allows researchers to prepare several common resolving-gel concentrations from the same stock solution, including 6%, 8%, 10%, 12% and 15% gels.
ProtoGel 40% for Western Blot Workflows
ProtoGel can be used to prepare SDS-PAGE gels for protein separation before Western blot transfer.
A typical workflow is :
Protein sample preparation → SDS-PAGE using ProtoGel → protein separation → membrane transfer → antibody detection
Because gel percentage influences protein migration and transfer performance, researchers should select the resolving-gel concentration according to their target protein molecular weight and downstream assay.
Why Use a Ready-to-Use Acrylamide/Bis-Acrylamide Solution?
Preparing acrylamide stock solutions from separate monomers adds weighing, mixing and preparation steps to an electrophoresis workflow.
A stabilized premixed solution such as ProtoGel 40% can help laboratories:
- Reduce reagent preparation time
- Reduce monomer-ratio calculation errors
- Improve consistency between gel batches
- Standardize SDS-PAGE workflows between users
- Quickly prepare different gel concentrations
- Maintain a consistent 37.5:1 acrylamide/bis-acrylamide ratio
This can be especially useful for laboratories routinely casting multiple SDS-PAGE gels.
ProtoGel 40% Product Specifications
| Specification | ProtoGel 40% |
| Product name | ProtoGel 40% / ProtoGel 0.4 |
| Catalog number | EC-891 |
| Manufacturer | National Diagnostics |
| Concentration | 40% |
| Acrylamide : Bis-acrylamide | 37.5:1 |
| Primary application | Protein electrophoresis |
| SDS-PAGE | Yes |
| Laemmli gels | Yes |
| Format | Stabilized ready-to-use solution |
| Common gel percentages | 6%, 8%, 10%, 12%, 15% |
| Sizes | 450 mL and 1 L |
ProtoGel 40% Frequently Asked Questions
What is ProtoGel 40%?
=> ProtoGel 40% is a stabilized acrylamide/bis-acrylamide stock solution with a 37.5:1 ratio used for preparing polyacrylamide gels for protein electrophoresis.
Is ProtoGel 0.4 the same as ProtoGel 40%?
=> The Maxanim ProtoGel 0.4 product uses catalog number EC-891, which corresponds to National Diagnostics ProtoGel 40%.
What is ProtoGel EC-891 used for?
=> EC-891 is primarily used to prepare polyacrylamide gels for SDS-PAGE and protein separation.
How much ProtoGel 40% do I need for a 10% gel?
=> For a final volume of 10 mL, a 10% gel requires 2.5 mL of ProtoGel 40%.
The calculation is:
10 × 10 ÷ 40 = 2.5 mL
How much ProtoGel 40% do I need for a 12% gel?
=> For a 10 mL resolving gel:
12 × 10 ÷ 40 = 3.0 mL ProtoGel 40%
What acrylamide-to-bisacrylamide ratio does ProtoGel use?
=> ProtoGel 40% uses a 37.5:1 acrylamide-to-bisacrylamide ratio.
Can ProtoGel be used for Western blotting?
=> Yes. ProtoGel-based SDS-PAGE gels can be used to separate proteins before transfer to a membrane for Western blot analysis.
Which buffer is used with ProtoGel?
=> A resolving buffer is used for the resolving gel and a stacking buffer for the stacking gel. Standard Laemmli SDS-PAGE can subsequently be run using Tris-Glycine-SDS running buffer.
Where can I order ProtoGel 40%?
=> ProtoGel 40% / ProtoGel 0.4, catalog number EC-891, can be requested through Maxanim. Availability, pack size, shipping and current pricing should be confirmed before ordering.
CTA: Request ProtoGel EC-891 Pricing & Availability
Safety Information
ProtoGel contains unpolymerized acrylamide and must be handled as a hazardous laboratory reagent by appropriately trained professional users.
Read the current Safety Data Sheet before use and follow your institution's procedures for personal protective equipment, handling, storage, spill response and hazardous-waste disposal.
Order ProtoGel 40% / EC-891
Looking for a consistent acrylamide/bis-acrylamide solution for SDS-PAGE gel preparation?
ProtoGel 40% (EC-891) provides researchers with a convenient 37.5:1 stabilized monomer solution for preparing a range of polyacrylamide gel concentrations.
Available formats: 450 mL and 1 L
- CTA : Check Availability
- CTA :Request a Quote
- CTA :Contact Technical Support