Nacalai USA, Inc. | Product | WIDE RANGE Gel Preparation Buffer (4x) for PAGE

WIDE RANGE Gel Preparation Buffer (4x) for PAGE

Simple casting procedures! Gradient gel-like separation / Hand-made Gel Preparation Buffer

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.

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.

Comparison of Separation Patterns with gradient gels

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

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.

Application: Detection of β-Actin by Western Blotting

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

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

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 (Cat. No.: 06865)

Recipe for WIDE RANGE Gel

The Acryamide/Bis premixed solutions used in this table are as follows;
40(w/v)%-Acrylamide/Bis mixed Solution (29:1) (Cat. No.: 16119)
40(w/v)%-Acrylamide/Bis mixed Solution (37.5:1) (Cat. 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	4.5	6
	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	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	7.5	10.0
	WIDE RANGE Gel Preparation Buffer	2.5	5	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	6.0	9	12
	WIDE RANGE Gel Preparation Buffer	2.5	5	7.5	10
	10(w/v)%-Ammonium Peroxodisulfate Solution	0.1	0.2	0.3	0.4
	TEMED	0.006	0.012	0.018	0.024

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