![]() ![]() Load directly on gelsno pre-heating is necessary. When used during SDS-PAGE, these protein ladder sets provide molecular weight markers ranging from 10250 kDa (unstained ladder) or 10210 kDa (pre-stained ladder). Compatible with chemiluminescent substrates and fluorescent secondary antibodies. Convenient and ready-to-use CLEARLY Protein Ladder sets are mixtures of 11 highly purified recombinant proteins. The ladder will allow you to monitor protein separation during electrophoresis and subsequently verify protein weight in your sample during later analysis. Western blotting: detection of the two unstained bands via the detection method used for the target protein. Western blotting (protein blotting or immunoblotting) is a rapid and sensitive assay for the detection and characterization of proteins.Western blot confirmation-detect the two unstained proteins (30 kDa and 80 kDa) using the detection system for the target protein.Fluorescent visualization-detect the 10 stained proteins using the 670 nm red laser or 700 nm channel of a fluorescence imager Use Precision Plus Protein WesternC Standards as a combination standard for western blotting and fluorescence detection in SDS-PAGE.The protein ladder 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.Ĭompare and view all other protein standards and ladders › Do not heat, dilute, add reducing agent before loading.Īpproximately 0.2~0.4 mg/ml of each protein in buffer (20 mM Trisphosphate pH 7.5 at 25☌), 2% SDS, 1 mM 2-Mercaptoethanol, 3.6 M Urea, and 15% (v/v) glycerol).ĥ μl of 10-180kDa Prestained Protein Ladder resolves 10 bands in 4-20% SDS-PAGE (Tris-glycine buffer) and after Western blotting to PVDF membrane.IBright Prestained Protein Ladder contains twelve recombinant proteins, ten (11 to 250 kDa) that are blue-stained and fluor-labeled for direct and near-IR fluorescent visualization and protein sizing, and two (30 kDa and 80 kDa) that are unstained and contain IgG binding sites to bind the primary and secondary antibodies used for chemiluminescent and fluorescent detection of the target protein. ![]() The ladder is supplied in gel loading buffer and is ready to use. The 10-180kDa Prestained Protein Ladder is designed for monitoring protein separated during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes(PVDF, nylon, or nitrocellulose) and for approximate sizing of proteins. 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). Keep on ice throughout the western blot process. We recommend blocking 35 non-fat dry milk, BSA, or normal serum for 1 hr at room temperature. The 10-180kDa Prestained Protein Ladder is a three-color protein standard with 10 pre-stained proteins covering a wide range molecular weights for 10 to 180 kDa. Increase the blocking incubation period and consider changing the blocking agent. 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. Apply more for thicker (>1.5 mm) or larger gel. Prestained Protein Ladder ab116028 is a three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa.This mixture can include all of the proteins associated with a particular tissue. 2~3 μl per well for general Western transfering. A western blot is a laboratory method used to detect specific protein molecules from among a mixture of proteins.3 μl or 5 μl per loading for clear visualization during electrophoresis on 15 well or 10 well mini-gel respectively. ![]()
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