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Invitrogen™ NuPAGE™ Large Protein Sensitive Staining Kit
Description
The HiMark™ Pre-Stained and Unstained High Molecular Weight Protein Standards offer a time-saving load-and-go format. The HiMark Pre-stained Standard includes one pink colored orientation band and eight blue bands ranging in apparent molecular weight of 31 - 460 kDa in NuPAGE Tris-Acetate Gel/SDS buffer system. It's suitable for monitoring electrophoresis, western transfer, and approximating molecular weight of unknown big proteins. The HiMark Unstained Standard, along with the NuPAGE Tris-Acetate Gel/SDS buffer system, offers the highest accuracy for molecular weight estimation of large proteins. It consists of nine proteins, ranging in size from 40 to 500 kDa in NuPAGE Novex 3-8% and 7% Tris-Acetate Gel/SDS buffer systems. Visualize with Coomassie™ stain, silver, and fluorescent protein stains post-electrophoresis. Several NuPAGE Large Protein Starter Kits are available to provide you with all necessary reagents and optimized protocols for high-resolution large protein separation and analysis. Each kit includes two boxes of NuPAGE 3-8% Tris-Acetate Gels, Tris-Acetate SDS Buffer Kit, a staining reagent or blotting membranes, and a vial of the appropriate HiMark Standard.
- Gel Percentage: 3-8 %
- Gel Size: Mini (8 x 8 cm)
- Well Format: 1D Well
1D Gel Electrophoresis, Protein Gel Electrophoresis, Protein Gel Staining and Imaging, Proteins, Expression, Isolation and Analysis
Specifications
Specifications
| Content And Storage | See product manual for kit contents and component storage conditions. Components of each kit are also available individually. |
| Sample Loading Volume | 25 μL |
| Size Range | 40 to 500 kDa |
| Quantity | 20 gels kit |
| Thickness | 1.0 mm |
| Well Design | 1D Well |
| Product Line | NuPAGE |
| Product Type | Protein Stain Kit |
| For Use With (Equipment) | XCell SureLock Mini-Cell |
| Stain Type | Unstained |
Frequently Asked Questions (FAQs)
DTT is not stable, so it must be added and the reduction performed just prior to loading your samples.
Precipitation of the LDS or SDS at 4 degrees C is normal. Bring the buffer to room temperature and mix until the LDS/SDS goes into solution. If you do not want to wait for it to dissolve, you can store the sample buffer at room temperature.
While they are both Bis-Tris based gels, the chemistries are very different since Bolt gels are optimized for western blotting. Another key difference is the wedge well design of the Bolt gels, which allows larger sample volumes to be loaded.
- Increase the pH of Tris-Glycine transfer buffer to 9.2, allowing all the proteins below pI 9.2 to transfer towards the anode electrode.
- Use the Tris-Glycine transfer buffer and place a membrane on both sides of the gel. If there are any proteins that are more basic than the pH of the transfer buffer, they will be captured on the extra membrane placed on the cathode side of the gel. Both membranes can then be developed in the same manner.
- Prior to blotting, incubate the gel for 15 minutes in Tris-Glycine transfer buffer containing 0.1% SDS. The small amount of SDS will give the proteins enough charge to move unidirectionally towards the anode and in most cases, should not denature the protein. Proceed with the transfer using regular Tris-Glycine transfer buffer.
For Research Use Only. Not for use in diagnostic procedures.