Two-Dimension Electrophoresis Gallery
This page shows two-dimension electrophoregrams of various tissues from various laboratory animals, obtained in our laboratory.
Please contact us if you have any questions and/or suggestions.
Methods in Two Dimension Electrophoresis
Sample preparation
Tissues/cells were suspended in protein extraction buffer (ReadyPrep™ protein extraction kit (Total Protein); Bio-rad), and homogenized with a bead-mill (TyssueLyser; Qiagen). After centrifugation, the supernatant was collected and its protein content was quantified with EZQ® Protein Quantification Kit (Invitrogen). The samples were reduced and alkylated with ReadyPrep™ reduction-alkylation kit(Bio-rad), and then applied for two-dimension electrophoresis.
First dimension: Isoelectic focusing
Samples were diluted with Rehydration/sample buffer (included in ReadyPrep™ 2-D starter kit; Bio-Rad), and used for isoelectric focusings (IEFs) with ReadyStrip™ IPG strips (pH3-10; Bio-rad) and PROTEAN® IEF cell(Bio-rad).
Second dimension: SDS-PAGE
After IEFs, the strips were treated with equilibrium buffers I and II (included in ReadyPrep™ 2-D starter kit; Bio-Rad), and then applied for SDS-PAGE. Precision Plus protein™ standard (unstained; Bio-rad) was used as molecular weight markers.
For SDS-PAGE, we used two gel systems:
- 10-20% gradient gel (ReadyGel J; Bio-rad) + Tris Glycine SDS buffer (Bio-rad)
- 4-12% gradient gel (NuPAGE® Novex BisTris Gel; Invitrogen) + MES buffer (Invitrogen)
Capture of gel images
Gels were stained with Sypro® Ruby gel stain (Invitrogen) and its images were captured with a laser scanner (FX-Pro; Bio-rad). The original images were converted into low-resolution jpegs with image software (Photoshop, Adobe) for easy handling on the web.
Gallery of 2DE images
Testis
AKR mouse
Sample: from an adult male (stored in RNAlater ®)
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
DBA/1 mouse
Sample: from an adult male (stored in RNAlater ®)
Sperm
Epididymal sperm from C57BL/6Cr mouse
Sample: from an adult male (directly put into Rehydration/sample buffer)
Ovary
Ovary from C57BL/6Cr mouse
Sample: from an 4-week-old female (stored in RNAlater ®, after buffer exchange using Zeba™ desalt spin column)
Ovary from C57BL/6Cr mouse (48 h after PMSG treatment)
Sample: from an 4-week-old female (stored in Allprotect Tissue reagent, Qiagen).
Protein was extracted using AllPrep DNA/RNA/Protein Mini Kit (Qiagen) and used without reduction-alkylation treatment before IEF.
Ovary from A/J mouse
Sample: from an 4-week-old female (stored in RNAlater ®, after buffer exchange using Zeba™ desalt spin column)
Ovary from DBA/1 mouse
Sample: from an 4-week-old female (stored in RNAlater ®, after buffer exchange using Zeba™ desalt spin column)
Ovary from DBA/2 mouse
Sample: from an 4-week-old female (stored in RNAlater ®, after buffer exchange using Zeba™ desalt spin column)
Ovary from DBA/2 mouse (48 h after PMSG treatment)
Sample: from an 4-week-old female (stored in Allprotect Tissue reagent, Qiagen).
Protein was extracted using AllPrep DNA/RNA/Protein Mini Kit (Qiagen) and used without reduction-alkylation treatment before IEF.
Ovary from C3H/He mouse
Sample: from an 4-week-old female (stored in RNAlater ®, after buffer exchange using Zeba™ desalt spin column)
Ovary from BDF1 mouse
Sample: from an 4-week-old female (stored in RNAlater ®, after buffer exchange using Zeba™ desalt spin column)
Ovary from BDF1 mouse (48 h after PMSG treatment)
Sample: from an 4-week-old female (stored in Allprotect Tissue reagent, Qiagen).
Protein was extracted using AllPrep DNA/RNA/Protein Mini Kit (Qiagen) and used without reduction-alkylation treatment before IEF.
Ovary from SD rat
Sample: from a non-pregnant adult female (stored in RNAlater ®; Its estrous stage was not monitored.)
Oocytes
Ovulated oocytes from BDF1 mice
Sample:Ovulated oocytes from BDF1 mice treated with eCG and hCG (directly put into Rehydration/sample buffer).
In vitro matured oocytes from BDF1 mice
Sample:In vitro matured oocytes from 21-day-old BDF1 mice (directly put into Rehydration/sample buffer).
In vitro matured oocytes from C57BL/6Cr mice
Sample:In vitro matured oocytes from 21-day-old C57BL/6Cr mice (directly put into Rehydration/sample buffer).
Molecular Standard: Mark12™ Wide Range Unstained Standard (Invitrogen).
In vitro matured oocytes from DBA/2 mice
Sample:In vitro matured oocytes from 21-day-old DBA/2 mice (directly put into Rehydration/sample buffer).
In vitro matured oocytes from ICR mice
Sample:In vitro matured oocytes from 21-day-old ICR mice (directly put into Rehydration/sample buffer).
Liver
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
AKR mouse
Sample: from an adult male (stored in RNAlater ®)
Heart
C57BL/6Cr mouse
Sample: from an adult male (stored at -80°C)
JW rabbit (Ventricle)
Sample: frozen tissue purchased from Funakoshi Co., Ltd.
Hartley guinea pig (Ventricle)
Sample: frozen tissue purchased from Funakoshi Co., Ltd.
SD rat (Ventricle)
Sample: frozen tissue purchased from Funakoshi Co., Ltd.
Syrian Hamster (Ventricle)
Sample: from an adult female (stored in RNAlater ®)
Skeletal muscle
C57BL/6Cr mouse, thigh muscle
Sample: from an adult male (stored at -80°C)
Molecular Standard: ECL Dual-vue Western Blotting Marker (GE Healthcare).
Kidney
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
Urine
BDF1 mouse
Sample: from an immature female, without reduction-alkylation treatment before IEF
Lung
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
Spleen
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
Thymus
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
Cerebrum
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
Cerebellum
C57BL/6J mouse
Sample: from an adult male (stored in RNAlater ®)
