![]() Storage Buffer: 62.5 mM Tris-H3PO4 (pH 7.5 at 25 ☌), 1 mM EDTA, 2% SDS, 10 mM DTT, 1 mM NaN3, 33% glycerol. Use 3uL to 5uL per well for in gel visualization, or 2.5uL per well for western blotting. Do not heat, dilute, add reducing agent before loading. My experimental protein size is 124kDA and I choose Beta Actin as a loading control which is small. The ladder is supplied in gel loading buffer and is ready to use. I am a beginner of western blot and from some days I have been facing a lot of problems. The Flash Protein Ladder is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins. Precision Plus Protein Unstained Recombinant Protein Standards are Strep-tagged, enabling immunodetection and molecular weight determination on western blots. Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Tris-glycine buffer). Unstained natural protein standards allow accurate MW determination with uniform band intensities on SDS-PAGE gels stained with Coomassie Blue or zinc. I am using freshly prepared buffers and solutions everytime but getting diffused bands in the gel (resolving gel - 10 and stacking gel - 5). The protein standard is supplied in a ready-to-use format for direct loading onto gels no need to heat, reduce, or add sample buffer prior to use. I am doing a western for my 75Kda protein of interest. The Flash Protein Ladder is a three-color protein standard with 13 pre-stained proteins covering a wide range molecular weights from 3.5 to 245 kDa. SeeBlue Plus2 Pre-Stained Standard contains 10 proteins (4250 kDa): 8 blue-dyed and 2 with contrasting colors, for easy and quick evaluation of electrophoresis and western transfer efficiency.
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