Western Blot Protein Standard

The MagicMark™ XP Western Protein Standard employs innovative technology for easy and accurate molecular weight estimation directly on western blots. It consists of a mixture of nine proteins ranging in mass from 20 to 220 kDa (Figure 1). Each protein band contains an IgG binding site, allowing marker visualization using the same reagents (chemiluminescent, fluorescent, or colorimetric substrates) and protocol for the target protein as your immunoassay. There are no extra procedures or additional reagents involved that can increase nonspecific background on your western blots (Table 1). In addition, the non–dye-modified proteins in the MagicMark™ XP Standard result in truer molecular weight estimations and sharper band resolution than with conventional protein standards.

Learn More About Novex® Protein Standards and Ladders
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Table 1. Comparison of various common protein molecular weight standards used on western blots.

Standard Type              Special RequirementsPotential ArtifactsHighly Accurate?
 Extra ReagentsAdditional Steps  
MagicMark™ XP Protein StandardNoneNone
  • None
Yes
Unstained Protein StandardsProtein Stain
  1. Cut standard lane
  2. Stain with protein stain
  3. Align, mark, and overlay with blot
  • Misaligned Bands
No
Prestained Protein StandardsNone
  1. Cut standard lane
  2. Align, mark, and overlay with blot
  • Mismarked Bands
No
Biotinylated or S-Protein StandardsConjugate (Avidin or S-Protein)
  1. Dilute standard
  2. Optimize conjugate dilution
  • Diffuse Bands
  • High Background
No

 

MagicMark XP Western Blot Protein Standard 


Figure 1. Sharp bands of MagicMark™ XP Western Protein Standard. Five-microliter samples of the MagicMark™ XP Western Protein Standard were run on each indicated gel type. Chemiluminescent detection was performed using the WesternBreeze® Anti-Rabbit Chemiluminescent Kit. Chromogenic detection was performed using the WesternBreeze® Anti-Rabbit Chromogenic Kit.
 

Properties of the Western Blot Protein Standard

ApplicationMagicMark™ XP Western Protein Standard
SDS-PAGEGood
Western BlotBest
Detection with/UV TransilluminationNA
Confirmation of Epitope-tagged ProteinsNA
Immediate Band IdentificationSee helpful tip for Western blot analysis below
Sharp BandsBest
MW EstimationBest
Monitor Migration during ElectrophoresisSee helpful tip for Western blot analysis below
Silver StainingGood
MW/pI Range20–200 kDa
IEF GelNA
Cat. No. LC5602

Helpful Tip

Combine MagicMark™ XP Standard and Novex® Sharp Pre-stained Standard, SeeBlue® Plus2 or SeeBlue® Pre-stained Standard or in the same run using the same lane. The colored bands from the pre-stained standards confirm gel run and transfer. Following immunodetection, sharp MagicMark™ XP bands develop directly on the western blot. Download a handy reference card on Western Protein Standards.