![]() ![]() For most researchers, we generally recommend:Ī) ab116027 for proteins between 10 and 180 kDaī) ab116028 for larger proteins up to 245 kDaĬ) ab116029 for smaller proteins down to 3.5 kDaĭ) ab116029 or ab234592 for studying smaller and larger proteins at the same time. When choosing the right pre-stained protein ladder, the most important choice is based on the size of your protein of interest. Prestained ladders also can use highlight bands of a different color, to make it easier to identify different size bands. That always affect the overall ubiquitination levels in the lyaste at a given time upon MG132 treatment. Protein ladders are most convenient to use when they are supplied ready to use, with no heating, dilution, or addition of reducing agent required before loading onto the gel. 1- Check what is the half life of your overexpressed protein-X. Block the membrane in TBS-T (TBS + 0.05 Tween-20) containing 5 milk powder for 1h at room temperature. Perform SDS-PAGE with a blue pre-stained protein molecular weight marker of your vendor of choice and your samples. It can be used with NuPAGE, tris-glycine, and tricine gels. Consisting of recombinant proteins fused to an IgG binding site, these versatile molecular weight ladders bind the primary or secondary antibody used for the detection of the target protein. Western Blotting Suggested dilution: 1:1,000 - 1:3,000 Western Blot protocol. The SeeBlue Plus2 Pre-Stained Standard contains 10 proteins: 8 blue-dyed and 2 with contrasting colors, providing easier band identification. Unstained protein ladders are more accurate for sizing proteins, as the dyes used in prestained ladders can slightly distort the apparent size of the protein ladder proteins on the gel. Western blot protein ladders are designed for protein molecular weight estimation directly on the blot during detection. However, an unstained protein ladder can only be visualized following staining with Coomassie or a similar non-specific protein stain. Using an unstained protein ladder is very useful when you need to accurately calculate the size of your protein. calculate the approximate size of your protein by overlaying an image of the membrane with the ladder with the image generated by antibody staining. A green and a red reference band, respectively, enable easier identification. Sizes range from 10-245 kDa suitable for accurate molecular weight determination of most cellular proteins. The BLUeye prestained protein ladder is provided in a gel loading buffer. Stable for up to 2 years if stored at -20☌ and supplied pre-stained in gel loading buffer for direct loading, Cleaver Scientific and BLUE Wide Range recombinant protein markers are perfect for SDS-PAGE applications. verify transfer efficiency between gel and membrane (PVDF, nylon, or nitrocellulose) The ready-to-use BLUeye prestained protein ladder has 12 sharp bands at 11, 17, 20, 25, 35,48, 63, 75, 100, 135, 180, and 245 kDa, which has more bands than commonly used markers. monitor protein separation during SDS-polyacrylamide gel electrophoresis Using a prestained protein ladder when running a western blot helps when you need to: ![]()
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