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Invitrogen™ NuPAGE™ Bis-Tris Welcome Pack, 12%
Description
The NuPAGE Bis-Tris Welcome Pack provides all of the necessary NuPAGE gels, buffers, and reagents you will need to begin using the Mini Gel Tank. The Mini Gel Tank is compatible with all Invitrogen Novex, NuPAGE, and Bolt mini gels. Each Mini Gel Tank can accommodate up to two gels per run. The unique tank design enables convenient, side-by-side gel loading and enhanced viewing during use. Run times may vary depending on buffer conditions and power supplies used.
The NuPAGE Bis-Tris Welcome Pack contains:
• Mini Gel Tank (A25977)
• NuPAGE Bis-Tris mini gels (2 boxes, 20 gels)
• NuPAGE MES SDS Running Buffer, 20X (NP0002)
• NuPAGE LDS Sample Buffer, 4X (NP0007)
• NuPAGE Sample Reducing Agent, 10X (NP0009)
• PageRuler Plus Prestained Protein Ladder, 10 to 250 kDa (26619)
About the Mini Gel Tank
The Mini Gel Tank is compatible with all Invitrogen Novex, NuPAGE, and Bolt mini gels. Each Mini Gel Tank can accommodate up to two gels per run. The unique tank design enables convenient, side-by-side gel loading and enhanced viewing during use. Run times may vary depending on buffer conditions and power supplies used.
About NuPAGE Bis-Tris gels
Invitrogen NuPAGE Bis-Tris Protein gels are precast polyacrylamide gels designed to give optimal separation of a wide range of proteins under denaturing conditions. Unlike traditional Tris-glycine gels, NuPAGE Bis-Tris gels have a neutral pH environment that minimizes protein modifications. Use NuPAGE Bis-Tris gels for preparing proteins for sequencing, mass spectrometry, and any other techniques where protein integrity is crucial. Also, use NuPAGE gels for optimal results during day-to-day use.
Transfer
For transferring your proteins to a membrane, we recommend using for NuPAGE Transfer Buffer (NP0006) for traditional wet transfer using the Mini Blot Module (B1000). Alternatively, rapid semi-dry transfer can be done using the Invitrogen Power Blotter or rapid dry transfer using the iBlot 2 Gel Transfer Device (IB21001).
Warranty
Up to 12 months
Specifications
Specifications
| Gel Percentage | 12% |
| Gel Size | Mini |
| Gel Thickness | 1.0 mm |
| Gel Type | Bis-Tris |
| Separation Range | 3.5 to 80 kDa |
| Wells | 15-well |
| Quantity | 1 Welcome Pk. kit |
| Shipping Condition | Approved for shipment at room temperature and wet ice |
| Product Line | NuPAGE |
| For Use With (Equipment) | Mini Gel Tank |
Frequently Asked Questions (FAQs)
- Decrease voltage, current or length of transfer time
- Make sure that the methanol concentration in the transfer buffer is proper; use a methanol concentration of 10-20% methanol removes the SDS from SDS-protein complexes and improves the binding of protein to the membrane.
- Make sure that the SDS concentration (if added) in the transfer buffer is proper, don't use more than 0.02-0.04% SDS. Using too much SDS can prevent binding of proteins to the membrane.
- Check the pore size of the membrane and the size of the target protein. Proteins smaller than 10 kDa will easily pass through a 0.45 µm pore size membrane. If proteins smaller than 10 kDa are of interest, it would be better to use a 0.2 µm pore size membrane.
- Increase voltage, current or length of time for transfer
- SDS in the gel and in the SDS-protein complexes promotes elution of the protein from the gels but inhibits binding of the protein to membranes. This inhibition is higher for nitrocellulose than for PVDF. For proteins that are difficult to elute from the gel such as large molecular weight proteins, a small amount of SDS may be added to the transfer buffer to improve transfer. We recommend pre-equilibrating the gel in 2X Transfer buffer (without methanol) containing 0.02-0.04% SDS for 10 minutes before assembling the sandwich and then transferring using 1X transfer buffer containing 10% methanol and 0.01%SDS.
- Methanol removes the SDS from SDS-protein complexes and improves the binding of protein to the membrane, but has some negative effects on the gel itself, leading to a decrease in transfer efficiency. It may cause a reduction in pore size, precipitation of some proteins, and some basic proteins to become positively charged or neutral. Make sure that the methanol concentration in the transfer buffer is not more than 10-20% and that high-quality, analytical grade methanol is used.
Pre-stained standards have a dye that is covalently bound to each protein that will result in the standard migrating differently in different buffer systems (i.e., different gels). As a result, using a pre-stained standard for molecular weight estimation will only give the apparent molecular weight of the protein. Pre-stained standards may be used for molecular weight approximation, confirming gel migration and estimating blotting efficiency but for accurate molecular weight estimation, an unstained standard should be used.
- While loading, take care to make sure that there is no cross-contamination from adjacent sample lanes.
- Make sure that the correct amount of standard is loaded per lane. Loading too much protein can result in extra bands and this is a problem especially with silver-stained gels.
- Improper storage of the standard or repeated freeze/thawing can result in protein degradation.
Here are some suggestions:
- Make sure that the correct amount of standard is loaded per lane. Loading too much protein can cause smearing and this is a problem especially with silver stained gels.
- Bands will not be as well resolved in low percentage gels. Try using a higher percentage gel.
- If the bands look smeary and non-distinct after a western transfer/detection, this may be due to the antibody being too concentrated. Follow the manufacturer's recommended dilution or determine the optimal antibody concentration by dot-blotting.