Simple casting procedures! Gradient gel-like separation

WIDE RANGE Gel Preparation Buffer (4x) for PAGE

Features

  • Simple casting procedures
    WIDE RANGE Gel Preparation buffer is a 4x concentrated neutral pH buffer. It can be used for preparation of both stacking gel and resolving gel by replacing the Tris-HCL buffer in Laemmli buffer system.
  • A wide separation range
    WIDE RANGE Gel provides a much greater separation range than the gel casted with a conventional Laemmli buffer system.
  • Improved stability and strength
    The increased tensile strength allows easy handling even a low percentage gel. The neutral pH buffer improves the stability of gel resulting in a longer shelf life than the gel with Laemmli buffer system.
Product Image

Gradient gels offer a much wider separation range of proteins than single percentage gels. However, casting gradient gels is more difficult and labor intensive. WIDE RANGE Gel Preparation Buffer offers a gradient gel-like separation on a single percentage gel by simply mixing it with acrylamide/ bisacrylamide gel casing solution. The gel can be used with the common sample buffer and running buffer. It is also suitable for standard staining methods including CBB and silver staining.

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Comparison of Separation

Gel size:

Sample:
Staining:
10 % WIDE RANGE gel and 12% Laemmli gel (10 cm x 8 cm x 1 mm)
Precast Gradient mini gels (dimensions vary by manufacturers)
Protein Marker (Product No. 29458)
CBB Stain One (Product No. 04543)
Handcast Single Percentage Gel Precast Gradient Gel

The results show that WIDE RANGE gel provides superior separation of the close size small proteins (6.5 KDa and 14 KDa) compared to the conventional Laemmli gel. Besides, the resolutions of high molecular weight proteins (45-200 KDa) are wider than Laemmli gel.

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Preparation of Different Percentage Gels with WIDE RANGE Gel Preparation Buffer

The Acryamide/Bis premixed solutions used in this table are as follows;
40(w/v)%-Acrylamide/Bis mixed Solution (29:1) (Product No. 16119)
40(w/v)%-Acrylamide/Bis mixed Solution (37.5:1) (Product No. 06121)

Preparation of Separation Gel (Unit: ml)

Acrylamide Reagent 10 ml 20 ml 30 ml 40 ml
6% Water 5.9 11.8 17.7 23.6
40(w/v)%-Acrylamide/Bis mixed Solution 1.5 3.0 4.5  6.0
WIDE RANGE Gel Preparation Buffer 2.5  5.0  7.5  10.0
10(w/v)%-Ammonium Peroxodisulfate Solution 0.1  0.2  0.3  0.4
TEMED 0.008 0.016 0.024 0.032
8% Water 5.4  10.8  16.2  21.6
40(w/v)%-Acrylamide/Bis mixed Solution 2.0  4.0  6.0  8.0
WIDE RANGE Gel Preparation Buffer 2.5  5.0  7.5  10.0
10(w/v)%-Ammonium Peroxodisulfate Solution 0.1  0.2  0.3  0.4
TEMED 0.006 0.012 0.018 0.024
10% Water 4.9 9.8 14.7 19.6
40(w/v)%-Acrylamide/Bis mixed Solution 2.5 5.0 7.5  10.0
WIDE RANGE Gel Preparation Buffer 2.5 5.0 7.5  10.0
10(w/v)%-Ammonium Peroxodisulfate Solution 0.1 0.2 0.3 0.4
TEMED 0.006 0.012 0.018 0.024
12% Water 4.4 8.8 13.2 17.6
40(w/v)%-Acrylamide/Bis mixed Solution 3.0 6.0  9.0 12.0
WIDE RANGE Gel Preparation Buffer 2.5 5.0 7.5 10.0
10(w/v)%-Ammonium Peroxodisulfate Solution 0.1 0.2 0.3 0.4
TEMED 0.006 0.012 0.018 0.024

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Preparation of Stacking Gel (Unit: ml)

Acrylamide Reagent 2 ml 4 ml 6 ml 8 ml
3% Water 1.33 2.66 3.99 5.32
40(w/v)%-Acrylamide/Bis mixed Solution 0.15 0.3 0.45 0.6
WIDE RANGE Gel Preparation Buffer 0.5 1 1.5 2
10(w/v)%-Ammonium Peroxodisulfate Solution 0.02 0.04 0.06 0.08
TEMED 0.002 0.004 0.006 0.008

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Gel Concentration

[Migration Table]

Gel Size:
Sample:
Staining:
10 cm x 8 cm × 1mm
Protein Marker (Product No. 29458)
CBB Stain One (Product No. 04543)

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Gel Strength

Even 6% wide range gel offers the same gel strength as 10% Laemmli gel.

Gel Strength

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Application: Detection of β-Actin by Western Blotting

WIDE RANGE gel can be used for western blotting with the standard detection protocol.

[Detection of β-Actin]

Gel

Sample

Blotting
Blocking
Primary Antibody
Secondly Antibody

Detection
10% WIDE RANGE Gel
12% Laemmli protocol gel
1 Chemi-Lumi One Markers(Product No. 06456)
2 Hela cell extract
PVDF membrane.................................................................................10V, 30min
Blocking One(Product No. 03953)......................................................60min
Mouse anti β-Actin, mono, 1:200.........................................................60min
Goat anti mouse IgG(H+L),conjugated HRP, 1:5000
+ Streptavidin(HRP Conjugate)(1:10000)(Product No. 06456)..... 60min
Detection Chemi-Lumi One Super(Product No. 02230)

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Application: 2D Electrophoresis with Silver Staining

WIDE RANGE gel is suitable for silver staining.

[Sample]
HL-60 cell extract
[Electrophoresis]
1D: Immobiline pH 4-7
2D: WIDE RANGE Gel buffer (12% gel)
[Staining]
Sil-Best Stain One(Product No. 06865)

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Protocol

References

  1. Y Chadani et al.: Proceedings of the National Academy of Sciences 113.7 (2016): E829-E838
  2. K Shcherbakova et al.: Genes to Cells 20.10 (2015): 789-801
  3. Y Yoshino et al.: Scientific reports 5 (2015)

Downloads

WIDE RANGE Gel Preparation Buffer (4x) for PAGEpdf (PDF 1 MB)

Ordering information

Product name Grade Storage Product No. PKG Size Online Catalog
WIDE RANGE Gel Preparation Buffer (4x) for PAGE SP R 07831-94 500 ml e-Nacalai

Related Products

Gel Casting Reagents
40(w/v)% - Acrylamide/Bis Mixed Solution (29:1) SP R 06119-45 500 ml e-Nacalai
40(w/v)%‐Acrylamide/Bis Mixed Solution (37.5:1) SP R 06121-95 500 ml e-Nacalai
30(w/v)%‐Acrylamide/Bis Mixed Solution (29:1) SP R 06141-35 500 ml e-Nacalai
30(w/v)%‐Acrylamide/Bis Mixed Solution (37.5:1) SP R 06144-05 500 ml e-Nacalai
10%‐SDS Solution [10%‐Sodium Lauryl Sulfate Solution] SP RT 30562-04 100 ml e-Nacalai
N,N,N',N'‐Tetramethylethylenediamine SP RT 33401-72 25 g e-Nacalai
10(w/v)%‐Ammonium Peroxodisulfate Solution SP F 02634-34 10 ml e-Nacalai
Running Buffer
Running Buffer Solution(10x) for SDS‐PAGE, Tris‐Glycine SP RT 30329-61 1 L e-Nacalai
Sample Buffer
Sample Buffer Solution with 2‐ME (2x) for SDS‐PAGE SP R 30566-22 25 ml e-Nacalai
Sample Buffer Solution without 2‐ME (2x) for SDS‐PAGE SP R 30567-12 25 ml e-Nacalai
Sample Buffer Solution with Reducing Reagent (6x) for SDS‐PAGE SP R 09499-14 5 ml e-Nacalai
Sample Buffer Solution without Reducing Reagent (6x) for SDS‐PAGE SP R 09500-64 5 ml e-Nacalai
Gel Steins
Dispotray (for minigel staining) RT 06563-44 20 Pieces e-Nacalai
Rapid Stain CBB Kit SP RT 30035-14 1 Set e-Nacalai
CBB Stain One (Ready To Use) SP RT 04543-51 1L e-Nacalai
Sil‐Best Stain One SP R 06865-81 1 Set e-Nacalai
Product Name Grade Storage Product No. PKG Size Online Catalog

(Storage) RT: Room temperature R: Refrigerator F: Freezer

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