SuperSep^™ Ace

	SuperSep™ Ace		SuperSep™ Ace Mini
Plate size	100 x 100 x 3 (mm)		100 x 60 x 4 (mm)
Gel size	90 x 85 x 1 (mm)		82 x 50 x 1 (mm)
Number of wells	13	17	17
Well Volume (μL)	30	25	20
Recommended electrophoresis chamber	EasySeparator™ (Product Number：292-36411)		EasySeparator™ Mini (Product Number：051-09251)

			Product Number
Proteins	Sample buffer (×2)	0.125M Tris-HCl, pH6.8、20% Glycerol、4% SDS、10% 2-Mercaptoethanol、0.004% BPB	196-11022
	Running buffer (×10)	0.25M Tris、1.92M Glycine、1% SDS	184-01291
Low molecular weight proteins	Sample buffer (×2)	0.125M Tris-HCl, pH6.8、20% Glycerol、4% SDS、10% 2-Mercaptoethanol、0.004% BPB	196-11022
	Running buffer (×10)	0.5M Tris、0.5M Tricine、1% SDS	200-17071
Nucleic acids	Sample buffer (×2)	0.01M Tris-HCl, pH7.9、1mM EDTA、30% Sucrose、0.004% BPB or Loading Buffer	313-90111
	Running buffer (×10)	0.25M Tris、1.92M Glycine or TBE buffer	318-90041

Features

• Good separation and clear bands (below)
• Can be used with multichannel pipette
• Patch on the side of the box to describe the gel type

Recommended electrophoresis conditions

• Constant current: 20 mA/gel
• Constant voltage: 200 V

Electrophoretic pattern by concentration

Select the acrylamide concentration based on the electrophoretic patterns presented below.

WIDE-VIEW™ Prestained Protein Size Marker III (Product Number: 230-02461) is used.

Examples of use

Example1 Electrophoresis of gel filtration-purified fractions for E. coli-expressed His-tagged protein

SuperSep™ was used for electrophoresis of gel filtration-purified fractions for E. coli-expressed His-tagged protein (35 kDa) to identify fractions containing the target protein.

Fraction No. 84 to 96 had a band at 35 kDa, showing that E. coli-expressed His-tagged protein was contained in Fractions No. 84 to 96.

Gel Running buffer Sample buffer Sample Molecular weight marker Stain

：SuperSep™ Ace 10-20%, 17 well (Product Number: 198-15041) ：SDS-PAGE Buffer, pH 8.5 (Product Number: 192-16801) ：Sample Buffer Solution (containing 3-mercapto-1, 2-propanediol) (×4) (Product Number: 196-16142) ：Gel filtration fractions for E. coli-expressed His-tagged protein (35 kDa) ：WIDE-VIEW™ Prestained Protein Size Marker III (Product Number: 230-02461) ：Quick-CBB PLUS (Product Number: 178-00551)

Example2 Determination of purified protein concentration

SuperSep™ was used for electrophoresis of E. coli-expressed His-tagged protein purified as described above in Example1 and albumin solutions to determine the concentration of purified protein.

The thickness of the bands detected showed that the concentration of purified E. coli-expressed His-tagged protein was approximately 0.45 mg/mL.

Gel Running buffer Sample buffer Sample Molecular weight marker Stain

：SuperSep™ Ace 10-20%, 17 well (Product Number: 198-15041) ：SDS-PAGE Buffer, pH 8.5 (Product Number: 192-16801) ：Sample Buffer Solution (containing 3-mercapto-1, 2-propanediol) (×4) (Product Number: 196-16142) ：E. coli-expressed His-tagged protein (purified protein) and 2 mg/mL albumin solution, bovine serum-derived (Product Number: 015-25613) ：WIDE-VIEW™ Prestained Protein Size Marker III (Product Number: 230-02461) ：Quick-CBB PLUS (Product Number: 178-00551)

Examples of use in native-PAGE

1. Analysis of the phospholipase C-δ1 pleckstrin homology domain using native polyacrylamide gel electrophoresis
   Analytical Biochemistry, Volume 431, Issue 2, 15 December 2012, Page 106–114
2. Core–shell clusters of human haemoglobin A and human serum albumin: artificial O2-carriers having various O2-affinities J. Mater. Chem. B, 2015, Advance Article
3. Bacterial toxin-inducible gene expression of cathelicidin-B1 in the chicken bursal lymphoma-derived cell line DT40: Functional characterization of cathelicidin-B1 Peptides, Volume 59, September 2014, Pages 94–102

Features

• Electrophoresis is completed in approximately 20 minutes (constant voltage: 300 V)
• Useful to confirm protein expression or purification
• High storage stability

Examples of use

SuperSep™ Ace Mini was used for electrophoresis of IgG antibody solutions. It was demonstrated that 20 minutes of electrophoresis resulted in good separation of markers and proteins in individual samples.。

Gel Running buffer Sample Molecular weight marker Stain Electrophoresis conditions

：SuperSep™ Ace Mini, 10-20%, 17 well (Product Number: 191-18613) ：SDS-PAGE Buffer, pH 8.5 (Product Number: 192-16801) ：Reduced/non-reduced IgG antibody solutions (+: heat-treated, -: non heat-treated) ：WIDE-VIEW™ Prestained Protein Size Marker III (Product Number: 230-02461) ：Quick-CBB PLUS (Product Number: 178-00551) ：Constant voltage (320 V), 20 minutes

CBB staining using a microwave

A secret technique for completing the entire process from electrophoresis using SuperSep™ Ace Mini^*1 to CBB staining to confirm protein expression in approximately 40 minutes^*2 is described below. Please try it.

• 1 SuperSep™ Ace can also be used.
• 2 The time required depends on the experimental conditions.

Reagents and instruments required

• Quick-CBB PLUS (Product Number: 174-00553)
• Microwave
• Plastic wrap
• Kimwipes^®
• Heat-resistant tray for holding gel

Procedure

1. SDS-PAGE
2. Staining using a microwave: 1 minute x 2 to 4 times
   Soak the gel in staining solution and then cover it with plastic wrap for 1-minute heating in a microwave (500 W). Repeat this procedure until the entire gel becomes colored.
3. Decolorization using a microwave: 1 minute x 4 to 6 times
   Discard the staining solution in the tray and pour 100 mL of deionized water to soak the gel. Heat the tray with rounded Kimwipes® inside in a microwave (500 W) for approximately 1 minute and then replace deionized water. Repeat the same procedure several times.^*3
   
   • 3 The gel under the solution is not broken by bumping. To remove the background, leave the gel in deionized water overnight.

Note:

： If the gel is completely covered with plastic wrap, make several holes with a needle. Since the tray becomes very hot, use gloves when removing it.

Staining results

• 

• 

• 
  
  
  

  Sample：BSA
  Amount of sample:
  ① 5 μg as total protein
  ② to ⑨ Two-fold serial dilution