Native PAGE

Reagents and Equipment

Sample Preparation for Native PAGE

SERVA Native PAGE Gels

SERVA Native PAGE Buffers

SERVA Protein Standards

SERVA Stains for Native PAGE

Native PAGE Equipment

 All you need for... Native PAGE

Blue and Clear Native electrophoresis in pH, the protein-dye complexes migrate pI-
polyacrylamide gels (BN/CN PAGE) sepa- independently towards the anode. The
rates proteins according to their native repulsion between the negatively charged
state, i.e. by their intrinsic charge and protein-dye complexes leads to high reso-
size. lution and band sharpness.
Blue Native PAGE (BN PAGE) makes use of Clear Native PAGE (CN PAGE) works with-
Coomassie® Brilliant Blue G 250 to bind to out using any anionic dye. Therefore,
the outer surface of protein complexes migration of proteins through the gel is as
leading to a negatively charged protein- well dependent from the intrinsic charge
dye complex. The Blue G dye does not act of the protein. This method can be used
as a detergent thus maintaining the native for separation of proteins with pI<7 at
structure of the protein throughout the physiological pH when dyes may interfere
electrophoresis process. At physiological with further analytical methods.

1 Sample Preparation for Native PAGE

2 SERVA Native PAGE Gels

3 SERVA Native PAGE Buffers

4 SERVA Protein Standards

5 SERVA Stains for Native PAGE

6 Native PAGE Equipment

SERVA produces gels for more than 30 years – hard to find a place with more experience
in manufacturing, developing and supporting the use of electrophoresis gels!

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1 Sample Preparation for Native PAGE

Detergents
To improve the solubility of hydrophobic ple preparations. They do not interfere
and membrane proteins you have to add with the electrophoretic run, but result in
non-ionic detergents to native PAGE sam- less streaking and better resolution.

Product Size Cat. No.

Digitonin 1g 19550.02
250 mg 19551.01
Digitonin water soluble
1g 19551.02
Dodecyl-beta-D-maltoside 1g 20780.03
100 ml 37242.01
Tergitol™ 15-S-9 500 ml 37242.02
2.5 L 37242.03

Enzymes
Cell and tissue lysates often have a high aration results.
DNA content, which causes a high viscosity Salt Active Nuclease is the choice for high
of samples. This impairs separation and salt samples. It digests DNA effectively
resolution of native PAGE. Salt Active Nucle- after dissociation of DNA-protein com-
ase is used for effective reduction of vis- plexes in high salt concentrations (up to
cosity caused by nucleic acids for best sep- 500 mM NaCl).

Product Size Cat. No.

Salt Active Nuclease 5.000 U 18541.01

0.25 M NaCl 0.5 M NaCl

SAN SAN SAN SAN

Incubation of E. coli lysate

2° C 37° C 2° C 37° C with increasing units of Salt
Active Nuclease for 30 min

Sample preparation is a crucial step in native PAGE 3

 2 SERVA Native PAGE Gels

SERVA developed precast gels opti- For native PAGE in a TRIS/Glycine

mized for a Blue Native/Clear Native buffer system SERVA offers the
PAGE system: SERVAGel™ N for BN SERVAGel™ PRiME™ gels showing
and CN PAGE, complemented by buff- highest resolution with long shelf life.
ers and Blue G solution for best results.

Lane 1 = Separation of stroma thylakoides mem- Separation of thylakoide membrane complexes

brane complexes by CN PAGE by 2D BN/SDS PAGE. By courtesy of Friedrich
Lane 2 = Separation of thylakoide membrane Ossenbuehl (University of Ulm).
complexes by BN PAGE. By courtesy of Friedrich
Ossenbuehl (University of Ulm).

Precast Gels for Native BN/CN PAGE 15 wells 12 wells 10 wells

SERVAGel™ N 3–12 % 43254.01 43250.01 43251.01
SERVAGel™ N 4–16 % 43255.01 43253.01 43252.01
SERVAGel™ N Native Starter Kit - - 43204.01

Precast TRIS/Glycine PRiME™ Gels 15 wells 12 wells 10 wells

SERVAGel™ TG PRiME™ 8 % 43284.01 43260.01 43261.01
SERVAGel™ TG PRiME™ 10 % 43285.01 43263.01 43264.01
SERVAGel™ TG PRiME™ 12 % 43286.01 43266.01 43267.01

SERVAGel™ TG PRiME™ 14 % 43287.01 43269.01 43270.01

SERVAGel™ TG PRiME™ 4–12 % 43288.01 43273.01 43274.01
SERVAGel™ TG PRiME™ 4–20 % 43289.01 43276.01 43277.01
SERVAGel™ TG PRiME™ 8–16 % 43290.01 43279.01 43280.01

4 Coming soon: SERVA horizontal large format gels for native PAGE.
3 SERVA Native PAGE Buffers
SERVA offers a complete range of buffers for native PAGE.

Product Size Cat. No.

Native Anode Buffer for Blue/Clear Native (10x) 1L 42535.01
Native Cathode Buffer for Blue/Clear Native (10x) 500 ml 42536.01

Sample Buffer for Blue Native (2x) 20 ml 42533.01

Sample Buffer for Clear Native (2x) 20 ml 42534.01
SERVA Blue G solution for BN, 1 % 20 ml 42538.01
SERVA Tris-Glycine Native Electrophoresis Buffer (10x) 1L 42530.01
SERVA Tris-Glycine Native Sample Buffer (2x) 20 ml 42528.01

4 SERVA Protein Standards

The SERVA Native Marker Liquid Mix The Protein Molecular Weight Stand-
for BN/CN PAGE is ready-to-use and ards Kit is a set of 8 single proteins. Pro-
contains 6 different proteins ranging teins are lyophilized and could easily be
from 21 up to 720 kDa. dissolved in water or sample buffer to
the desired final concentration.

Ferritin horse (Mr 450 000/720 000)

Urease jack bean (Mr 272 000/545 000)

Lactate dehydrogenase porcine (Mr 146 000)

Albumin bovine (Mr 67 000)

Albumin egg (Mr 45 000)

SERVA Native Marker Liquid Mix for BN/CN PAGE Trypsin inhibitor soybean (Mr 21 000)
(cat. no. 39219) separated by Clear Native PAGE
on SERVAGel™ N 4–16 % (cat. no. 43252)

Product Size Cat. No.

SERVA Native Marker Liquid Mix for BN/CN PAGE 5x 50 µl 39219.01

Protein Molecular Weight Standards* 1 kit 39064.01

*25 mg each of Ferritin horse, Mr 450 000; Catalase bovine, Mr 240 000; Aldolase rabbit, Mr 160 000;
Albumin bovine, Mr 67 000; Albumin egg, Mr 45 000; Chymotrypsinogen A, Mr 25 000; Myoglobin
equine, Mr 17 800; Cytochrome C, Mr 12 400.

Protein Standards in native PAGE are valuable tools to verify separation results. 5 5
 5 SERVA Stains for Native PAGE
Native PAGE gels are stained with buffer containing Coomassie ® Blue G
Commassie ®, silver or zinc-imidazole after half of the run time against cath-
stains. To receive best results for Blue ode buffer without dye.
Native PAGE gels change the cathode

Quick Coomassie® Stain Silver Staining Kit Native PAGE

1-step, non-toxic colloidal

Type of Stain Fast, MS-compatible silver stain
Coomassie ® G-250
Detection Visual, colorimetric Visual, colorimetric
Sensitivity 5 ng 1 ng
Staining time 15 min 45 – 60 min
Quantification  
MS compatible  
Re-usable  
No. of gels stained 40 - 120 25
Size 1L 1 kit
Cat. no. 35081.01 35077.01

Separation of increasing amounts of thylakoide membrane

from Synechocystis (lane 1 = 1 µg, … lane 10 = 10 µg). After
half of the run the blue cathode buffer has been replaced by
colourless cathode buffer. By courtesy of Friedrich Ossen-
buehl (University of Ulm).

SERVA offers a broad range of staining kits for protein gels, not only for
native PAGE but also for SDS PAGE, IEF and 2D electrophoresis.
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6 Native PAGE Equipment

BlueVertical™ PRiME™ & TankBlotter

▐▐ Dual mini tank systems
▐▐ Accommodates 1 – 2 gels in cassettes with outer dimensions
of 10 cm x 10 cm x 0.7 cm
▐▐ Leak-free inner core unit with a unique clamp system
▐▐ Unique, easy-to-handle blotting insert for tank blotting
▐▐ Gel Casting Stand for pouring two gels
▐▐ Quality designed and made in Germany

Product Size Cat. no.

BlueVertical™ PRiME™ 1 unit BV-104
BlueVertical™ PRiME™ TankBlot 1 unit BV-104-TB
BlueVertical™ PRiME™ Casting Stand 1 unit BV-104-CS
BlueVertical™ PRiME™ Blot Module 1 unit BV-104-B

BluePowerTM Power Supply

▐▐ For vertical mini gel electrophoresis: BluePowerTM 600 PRiMETM
Power Supply (600 V, 1000 mA, 300 W)
▐▐ Fully programmable (9 programms with 9 steps each)
▐▐ 4x 2 outlets

Product Size Cat. no.

BluePower TM 600 PRiME TM Power Supply 1 unit BP-600-PRI

Digital Imaging and Analysis System

▐▐ Fast and convenient gel documentation system
▐▐ Inlcudes darkroom cabinet, UV filter, digital SLR camera
▐▐ Optional available: UV-, white- and blue-light transilluminator,
epi-white light, LabImage 1D analysis software
▐▐ Stable metal housing
▐▐ Large door for easy gel handling
▐▐ WxHxD: 420 mm x 550 mm x 520 mm; weight: approx. 12 kg

Product Size Cat. no.

Digital Imaging and Analysis System III 1 system DIAS-III

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SERVA Electrophoresis GmbH

Carl-Benz-Str. 7
69115 Heidelberg / Germany
Fon: +49 6221 13840-0
Fax: +49 6221 13840-10
E-mail: info@serva.de · www.serva.de

Version 22/05