The apparent molecular weight of each protein (kDa) has been determined by calibration against an unstained protein ladder in each electrophoresis condition. Tested in polyacrylamide gel electrophoresis and Western Blotting. One prominent feature of this protein ladder, is that it remains in the liquid state at the storage temperature therefore, there is not a freeze-thaw cycle involved, making the ladder very stable.Ġ.1~0.4 mg/ml of each protein in 20 mM Trisphosphate (pH 7.5 at 25☌), 2% SDS, 1 mM reducing reagent, 3.6 M Urea, and stabilizer.įor each application, 5-15µl is needed, depending on the size of the wells. No need to heat, dilute, or add reducing agent before loading. The ladder is supplied in protein gel loading buffer and is ready-to-use. Two orange reference bands at 70 kDa and 25 kDa and one green reference band at 10 kDa highlight the blue-stained protein ladder. It is designed for monitoring protein separated during polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes and for sizing of proteins. The PageRuler Plus Prestained Protein Ladder is a mixture of nine (9) blue-, orange- and green-stained proteins (10 to 250 kDa) for use in protein electrophoresis (SDS-PAGE) and western blotting. The Bullseye Pre-stained Protein Ladder contains 10 prestained proteins, covering a wide range of molecular weights from 10 to 180kDa. During electrophoresis, the polypeptides of the ladder show up in the gel as distinctly colored bands. What this means is that before even loading the ladders into the gel for electrophoresis, these ladders are previously stained and ready to go. Western Blotting Reagents 2.4 NF-kB Reporter Assay Using RAW-Blue Cells. In a prestained protein ladder, each of these bands, or polypeptides, are already stained with a dye. Use 5-15µl, depending on the size of the wellsĪpplication: The Bullseye Pre-stained Protein Ladder is suitable for visualizing proteins during electrophoresis without staining and for monitoring transfer onto membrane. protein concentrations were determined using Pierce BCA protein analysis kit.2 reference bands - red at ~75kDa and green at ~25kDa.Load ladder into SDS-PAGE gel or Western blot using these volumes (based on gel thickness of 0.75 1.0mm). Mix solution gently to ensure that it is homogeneous. Stoolfix Gut Microbiome Stabilization Kitsīullseye Pre-stained Protein Ladder, 10-180 kDa mPAGE ® Western Protein Standards consist of seven recombinant proteins at 20 kDa, 30 kDa, 40 kDa, 50 kDa, 60 kDa, 80 kDa and 120 kDa with an IgG binding site that binds to most antibodies. Thaw ladder at room temperature to dissolve precipitated solids.Beakers, Graduated Cylinders, and Flasks.Aluminum Foil, Parafilm, and Multiwipes 5l of IRIS11 Prestained Protein Ladder (PMI11) resolves 11 bands in 4-20 SDS-PAGE (Tris-glycine buffer) and after Western blotting to PVDF membrane.Thaw the ladder either at room temperature or at 37-40☌ for a few minutes to dissolve precipitated solids. The quality of the BLUeye Prestained Protein Ladder is tested on a lot-to-lot basis to ensure consistent product quality.īLUeye Prestained Protein Ladder Protocolġ. ![]() Sizing of proteins on SDS-PAGE and Western blots.Īpproximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Trisphosphate, pH 7.5 at 25☌), 2 % SDS, 0.2 mM Dithiothreitol, 3.6 M Urea, and 15 % (v/v) Glycerol. Monitoring of protein transfer onto membranes during Western blots. ![]() Monitoring of protein migration during SDS-PAGE. These protein standards and ladders are consistent from lot to lot and are strictly. Each protein standard or ladder is supplied in a ready-to-use format, eliminating the need to reduce, pre-mix, or add loading dyes. Ready-to-use: supplied in a loading buffer for direct loading on gelsĮasy to identify: includes the ~25, ~75 kDa reference bands coupled with a green and a red dye Choose from a variety of protein ladders (molecular weight markers) for protein electrophoresis and western blotting applications. Stable at -20C for 2 years Stable at 4C for 6 months. The ladder is supplied in gel loading buffer and is ready to use.īroad range: 10-245 kDa (Tris-glycine-SDS running buffer) Quality Control Tested in SDS-polyacrylamide gel electrophoresis and western blotting. The BLUeye Prestained Protein Ladder is designed for monitoring protein separation during, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size 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-polyacrylamide gel electrophoresis (SDS-PAGE) with Tris-glycine-SDS running buffer. The BLUeye Prestained Protein Ladder is a three-color protein standard with 12 prestained proteins covering a wide range molecular weights from 10 to 245 kilodalton (kDa).
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