Western blot no protein or ladder4/11/2024 The rest of the proteins on your gel will of course be invisible, but you can easily see if the brightly colored ladder bands are still in the gel, or have transferred completely to the membrane. Use a pre-stained protein ladder to track transfer of proteins from a gel to a membrane. So how do you know when your transfer is complete? Here are a few good controls you can use to monitor transfer efficiency: Look for complete transfer of the molecular weight ladder Instead, you will need to optimize the transfer time and conditions for yourself, based on the equipment you use and the proteins you are working with. Transfer times are empirical and based on your own particular samples, which means that there is no easy way to determine how long you need to run the transfer in order to get complete transfer of all your proteins. Now it’s time for the all-important transfer step, that tricky point that will determine the quality of your Western blot. NOT FOR USE IN DIAGNOSTIC PROCEDURES (EXCEPT AS SPECIFICALLY NOTED).So, you’ve done your experiment, prepped your samples, and run your SDS-PAGE gel. Our mission is to develop high-quality innovative tools and services to accelerate discovery.įOR RESEARCH USE ONLY. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. Certificates of Analysis and Kit Components Lists are located under the Documents tab. Please see the Kit Components List to determine kit components. Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. CLEARLY Stained Protein Ladder is sufficient for 50 lanes. QuantityĬLEARLY Protein Ladder (Unstained) is sufficient for 100 lanes. They remain stable for up to 50 freeze-thaw cycles. These products are stable for 6 months when stored at 4☌. Ensure that the ladder is completely thawed before removing an aliquot for loading.Pre-stained ladder (Cat.# 3454): Load 10 µl per lane.Unstained ladder (Cat.# 3453): Load 5 µl per lane.*Reference band (four-fold greater intensity relative to the other bands) SDS-PAGE recommendations CLEARLY Stained Protein Ladder (Cat.# 3454A and 3454B).*Reference band (four-fold greater intensity relative to the other bands) CLEARLY Protein Ladder (Unstained) (Cat.# 3453A and 3453B).# 3453A for complete product documentation and resources. In this case, this ladder is sufficient for 100 lanes. Cat. 5 μl of this ladder is loaded per lane of SDS-PAGE minigel for standard use. This ladder is pre-mixed with loading buffer and ready to load. The 30 and 80 kDa bands have quadruple the mass of the other bands and serve as reference indicators. This ladder is intended for use as a precise size standard when performing SDS-PAGE to calculate the molecular weight of a protein of interest. When this ladder is analyzed by SDS-PAGE and stained with Coomassie Brilliant Blue R-250, it is resolved into quite clearly identifiable bands from 10 to 250 kDa. Migration pattern of CLEARLY Stained Protein Ladder (Cat.# 3454A) on an 12% SDS-PAGE gel Migration pattern of CLEARLY Stained Protein Ladder (Cat.# 3454A) on an 12% SDS-PAGE gel.ĬLEARLY Protein Ladder (Unstained) is a mixture of 11 recombinant highly purified proteins. *: Because the covalent coupling of the dye to the proteins affects their electrophoretic behavior on SDS-PAGE gels relative to unstained proteins, this ladder is not recommended for use as a precise size standard when performing SDS-PAGE to calculate the molecular weight of a protein of interest. # 3454A for complete product documentation and resources. *: Because the covalent coupling of the dye to the proteins affects their electrophoretic behavior on SDS-PAGE gels relative to unstained proteins, this ladder is not recommended for use as a precise size standard when performing SDS-PAGE to calculate the molecular weight of a protein of interest. Cat. This ladder is useful for monitoring of protein migration during SDS-PAGE and verifying transfer efficiency at western blotting. In this case, this ladder is sufficient for 50 lanes. 10 μl of this ladder is loaded per lane of SDS-PAGE minigel for standard use. The 30 and 70 kDa bands have quadruple the mass of the other bands and serve as reference indicators. When this ladder is analyzed by SDS-PAGE, it is resolved into clearly identifiable visible bands from 10 to 210 kDa. CLEARLY Stained Protein Ladder is a mixture of 11 pre-stained recombinant highly purified proteins.
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