Perfect Protein™ Markers
Precisely sized, conveniently spaced for accurate protein size determination
The Perfect Protein™ Markers are a novel set of recombinant proteins with defined sizes at convenient intervals. Designed for routine use in SDS-polyacrylamide gel electrophoresis, the Perfect Protein Markers enable highly accurate size determination of unknown samples. Unlike many conventional markers (e.g., ovalbumin, serum albumin, etc.), the Perfect Protein Markers contain no oligosaccharides that cause anomalous migration, heterogeneous “fuzzy” bands, or inaccurate size estimation. The known mass of each Perfect Protein Marker band also enables estimation of concentration of sample proteins. The markers are optimized for use with Coomassie blue staining, but adjusted amounts can also be used with other gel staining methods (e.g., silver staining, fluorescent dyes, etc.).

The Perfect Protein Markers, 15–150 kDa, include protein sizes of 15, 25, 35, 50, 75, 100, and 150 kDa. Each vial contains 400 mg protein (50 mg per band except for 100 mg of the 50-kDa high-intensity reference band). The Perfect Protein Markers, 10–225 kDa, include the protein sizes listed above and two additional proteins, 10 kDa and 225 kDa, for applications requiring a broader size range. Each vial contains 500 mg protein (50 mg per band except for 100 mg of the 50-kDa high-intensity reference band). The 4X SDS Sample Buffer is a standard formulation commonly used for SDS-PAGE analysis of proteins. The solution includes DTT for complete denaturation of disulfide bonds. The buffer can be used at a final concentration of 2X for most applications.

Perfect Protein™ Markers, 15–150 kDa
100 lanes
69149-3
Perfect Protein Markers, 10–225 kDa
100 lanes
69079-3

Perfect Protein™ Western Markers and Blot Kits
Accurate size markers detectable on any Western blot
The Perfect Protein™ Western Markers consist of seven recombinant S•Tag™/His•TagŪ fusion proteins that can be detected on any Western blot using S-protein AP or HRP Conjugates (or the His•Tag Monoclonal Antibody with AP-conjugated secondary antibody). These markers serve as precise size standards that appear simultaneously on the blot with target proteins, eliminating the uncertainty and imprecision associated with other methods. The Perfect Protein Western Blot Kits include the markers plus either S-protein AP or HRP Conjugate. For a version containing prestained proteins for tracking electrophoresis and Western transfer, please see Trail Mix™ Western Markers and Blot Kits in this chapter.

Features

• Detect markers simply by adding the S-protein (AP or HRP Conjugate) or His•Tag Monoclonal Antibody to the same incubation used for sample detection (Anti-Mouse IgG AP Conjugate or HRP Conjugate).
• S-protein conjugate (included in kits) does not interfere with antibodies or streptavidin detection.
• Can be used with colorimetric and chemiluminescent AP or HRP substrates.
• Recombinant, unmodified markers give sharp, accurately sized bands.
• Markers are supplied at the working dilution in gel loading buffer; concentration optimized for Western detection.
• Protein sizes are 15, 25, 35, 50, 75, 100, and 150 kDa.

Perfect Protein™ Western Markers
25 lanes
69959-3
Perfect Protein AP Western Blot Kit
25 blots
69965-3
Perfect Protein HRP Western Blot Kit
25 blots
69078-3

Components for AP and HRP Kits:
• 25 lanes Perfect Protein Western Markers
• 50 ml S-protein AP or HRP Conjugate

A. Two parallel blots were incubated with primary antibodies for an expressed protein, then incubated with Anti-Mouse IgG AP Conjugate and processed using chemiluminescent detection. The S-protein AP Conjugate was included with secondary antibody for the blot on the left. Lanes: 1, 5 ml Perfect Protein Western Markers; 2, 10 ng induced cell extract; 3, 1 mg uninduced cell extract.
B. Blot contains the Perfect Protein Western Markers plus purified GUS expressed in insect cells incubated with a GUS-specific monoclonal antibody, then incubated with Anti-Mouse IgG HRP Conjugate and S-protein HRP Conjugate and detected with chemiluminescent HRP substrate. Lanes: 1, 5 ml Perfect Protein Western Markers; 2, 800 ng purified GUS protein.

Trail Mix™ Protein Markers
Novel protein markers for visible tracking and accurate sizing in stained gels
Trail Mix™ Protein Markers are a mixture of the Novagen Perfect Protein™ Markers supplemented with a group of three prestained indicator proteins to allow direct visualization of protein migration during electrophoresis. Unlike other marker sets in which the entire ladder is prestained, Trail Mix uses only three reference bands (at 100, 16, and 15 kDa) to confirm separation and indicate gel orientation. Prestaining can cause band broadening or affect mobility, reducing the precision with which mobility and molecular weight determinations can be made. The migration or band sharpness of the Perfect Protein Markers is unaffected by the presence of the prestained bands in Trail Mix.
When stained with Coomassie blue, 10 bands appear, ranging from 10 kDa to 225 kDa. In addition to the prestained bands, the 50 kDa marker serves as a landmark on stained gels because it is present at a higher concentration in the mixture relative to the adjacent bands. The 4X SDS Sample Buffer is a standard formulation commonly used for SDS-PAGE analysis of proteins. The solution includes DTT for complete denaturation of disulfide bonds. The buffer can be used at a final concentration of 2X for most applications.
Trail Mix™ Protein Markers
100 lanes
70980-3

Trail Mix™ Western Markers and Blot Kits
Novel protein markers for visible tracking and accurate sizing on any Western blot
Trail Mix™ Western Markers consist of the Perfect Protein™ Western Markers supplemented with a group of three prestained indicator proteins to allow direct visualization of protein mobility during electrophoresis. Trail Mix Western Markers and Western Blot Kits have all of the features of Perfect Protein Western Markers and Western Blot Kits, plus easy to track prestained bands at 15, 16, and 100 kDa. This mix also contains an additional Perfect Protein Western Marker at 225 kDa for improved sizing accuracy of very large proteins.

Features

• Trail Mix contains three prestained indicator proteins plus eight unstained Perfect Protein Western Markers.
• Perfect Protein Western Marker concentrations are optimized for blot detection.
• Each Perfect Protein Western Marker carries an S•Tag™ and His•TagŪ fusion peptide.
• Kits contain S-protein AP or HRP Conjugate for convenient Western blot detection of markers.
• Conjugates can be added together with secondary antibody or streptavidin conjugates for simultaneous detection of target proteins and markers.
• Markers are supplied at the working dilution in gel loading buffer.
• Prestained proteins migrate at 15, 16, and 100 kDa. Protein sizes detected on Western blots are 15, 25, 35, 50, 75, 100, 150, and 225 kDa.