Equipment:

Instrument

PrimeApplications

R ehydration Chamber (Cytiva)

Iso-electric focusing (IEF) strip Rehydration

• Rehydrates (uptake of protein solution) up to 12 IPG strips, in lengths up to 24 cm. But 13 cm Strip will be preferred for the consistency of 2D-GE experiment.

• The chamber is stable against chemicals.

• Binding capacity to protein is very low.

EttanPHOR IPG (Cytiva)

Iso-electric focusing (IEF)

• Ettan IPGphor 3 IEF system is a fully integrated isoelectric focusing system optimized to deliver speed and reliability in the first dimension (Iso-electric focusing) of 2-D electrophoresis.

• Run up to 12 IPG strips (7, 11 and 13 cm) simultaneously using Ettan IPGphor manifold, Ettan IPGphor Manifold Light, or ceramic strip holders.

• Lid design allows optimum running with light-sensitive protein samples.

TE 22 Mini Tank Transfer Unit (Cytiva)

Protein transfer for blotting

• TE 22 Mini Tank Transfer Unit has been designed to deliver outstanding mini-gel electrophoretic tank transfer for western, southern and northern blots.

• For 9 x 10 cm of four mini-gels in about 1 litter of buffer the transfers can be completed in less than 1 hour.

• It has the built-in alumina-covered heat exchanger and tap water running through the system which dissipates the heat generated by tank electron transfer. The temperature rises no more than 5°C during a typical transfer.

2 D Ruby apparatus (Cytiva)

Run denaturing second dimension electrophoresis of proteins on acrylamide gels

• Temperature 1ºC to 45ºC with built in heat exchanger.

• Use two 7cm IPG strips side-by-side, allowing eight second-dimension separations at once.

• Two 13 cm strips can also be used.

Amersham Imager 680 (Cytiva)

Amersham Imager 680 is a blot and gel imager for sensitive, publication-quality imaging of DNA and protein gels. It has both chemiluminescence and fluorescence imaging facilities.

Illumination options in Amersham Imager 680

Chemiluminescence: Use with any blot that incorporates chemiluminescence labeling.

Colorimetric Epi-White: Take real-color images and use this with chemiluminescence mode to generate images for gel documentation or combine chemiluminescence with automatic color-marker overlay for easy molecular weight identification.

Colorimetric Trans-White: Factory calibrated system for quantitative OD measurement for Coomassie Brilliant Blue stained and silver stained gels.

Fluorescence Trans-UV: Use with ethidium bromide or other UV fluorescent stains.

Fluorescence Epi-RGB: Use for multiplex, quantitative Western blotting applications.

Amersham Typhoon Scanner (Cytiva)

• Amersham Typhoon scanners are designed for versatile imaging and precise quantitation of fluorescent, colorimetric, and radiolabeled proteins.

• The scanner has three different excitation wavelengths such as blue, green and red for Cy2 (488 nm), Cy3 (532 nm) and Cy5 (633 nm) dyes, respectively.

• These are mainly useful for 2-D fluorescence Difference Gel Electrophoresis (2-D DIGE) to quantify proteins from different subjects or phenotypes.

• The scanner also has NIR wavelengths (685 nm and 785 nm).

• The differential expression of proteins can be analyzed by DeCyder software.

DeCyder Software (Cytiva)

Data analysis

• The DeCyder image analysis software is a standard for the simultaneous analysis of thousands of proteins across different groups to reveal differentially regulated proteins using 2D-DIGE technology.

• It has a powerful algorithm to analyze large data sets.