The Panorama™
Mouse/Rat Tissue Extract Protein Array Kit is a convenient and robust tool
for accurate determination of protein distribution and expression. The
array contains 62 extracts from 10 mouse and 12 rat tissues spotted on a
nitrocellulose-coated slide, and is a reliable platform for rapid
evaluation of protein expression.
Introduction Sigma's Panorama™
Mouse/Rat Tissue Extract Protein Array Kit (Product Code MRPA1) contains
62 extracts from 10 mouse and 12 rat tissues spotted on a
nitrocellulose-coated slide. Each tissue is represented by three extracts
(total tissue, nuclear and cytoplasmic cell fractions). All extracts were
prepared in denaturing, reducing buffer and boiled. Extracts of the same
origin (i.e., cytoplasmic) from different tissues were spotted at equal
protein concentrations. In addition, a series of positive and negative
controls were spotted. Information on specific positioning of extracts and
controls can be viewed at: www.sigma-aldrich.com/abarray.
Figure 1. Rabbit primary antibodies
to different protein targets were tested, each on a separate
slide: 1) negative control-only secondary antibody: anti-rabbit
IgG, peroxidase conjugate; 2) anti-androgen receptor (product code
A 9853); 3) anti-cofilin (product code C 8736); 4) anti-binculin
(product code V 4139) and 5) anti-neurofilament 200 (product code
N 4142). The antibodies were developed with anti-rabbit IgG,
peroxidase conjugate and a chemiluminescent substrate. Note the
different protein profile expression of the proteins.
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Experimental Conditions and Results Differential expression of proteins
in tissues. The Panorama™ Mouse/Rat Tissue Extract Protein Array Kit
was used to profile the expression of proteins in mouse and rat tissues
using antibodies to several protein targets. A labeling system with
secondary antibody conjugated to peroxidase and CPS1 chemiluminescent
substrate (product code CPS1) was used. Resulting chemiluminescence was
detected using autoradiograph film. As expected, each antibody showed a
different pattern of protein expression (Figure 1). The results clearly
show that some of the proteins are broadly expressed in many tissues while
others are more specific to certain tissues.
Figure 2. A comparison of
fluorescent and chemiluminescent labeling and detection methods.
Anti-neurofilament 200 (product code N 4142) was applied on two
slides. Slide A was further incubated with a peroxidase-conjugated
secondary antibody and developed with a chemiluminescent substrate
(product xode CPS1), while slide B was incubated with a Cy3
dye-conjugated secondary antibody. Similar results were obtained
with both detection methods.
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Comparison of fluorescent and chemiluminescent detection systems. Two
detection systems, fluorescent and chemiluminescent, were tested and
compared for sensitivity. A rabbit polyclonal antibody to neurofilament
protein was used in both assays. One slide was further incubated with goat
anti-rabbit IgG conjugated to Cy3, and the other slide with the same
secondary antibody was conjugated to peroxidase. Very similar results were
obtained, leading to the conclusion that, under the chosen conditions,
both methods are comparable (Figure 2).
Conclusion Panorama Mouse/Rat
Tissue Extract Protein Array kit provides a reliable platform for the
rapid evaluation of serum, hybridoma supernatant, ascites fluid, purified
antibody or fractionated sera. The mouse and rat tissue proteins spotted
are extracts of total tissue, nuclear and cytoplasmic cell fractions. The
array is suitable for detection of proteins in various tissues and
compatible with fluorescent or chemiluminescent detection systems.
Reference Correspondence and
requests for additional information should be addressed to: Eliezer Kopf
at ekopf@sial.com
Sigma-Aldrich Corporation 3050
Spruce St.
St. Louis, MO 63103 USA
Tel. 800-521-8956; Fax 314-771-5757
www.sigma-aldrich.com/
