#Protein ladder for western blot verification
#Protein ladder for western blot plus

No conjugate is needed for fluorescent blot detection. Western blotting protein standards can be used for both fluorescent visualization and colormetric or chemiluminescent immunodetection on western blots. Loading buffer: 30% (w/v) glycerol, 2% SDS, 62.5 mM Tris, pH 6.8, 50 mM DTT, 5 mM EDTA, 0.02% NaN 3, 0.01% bromophenol blue.įluorescent detection: prepare a 1:10 dilution of the WesternC Standard and load 5 µl per well. Packaging Options Number of Applications (Volume)

Fluorescent blot detection of prestained bands, with blue bands visualized in the red wavelength range and pink bands in the green wavelength range.

#Protein ladder for western blot verification

This 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. With a range of fluorescent dyes and antibodies for western blot. Multiplex fluorescent western blotting provides accurate, quantitative results, stable signals, and the ability to clearly evaluate multiple protein targets on a single blot, which makes this technique increasingly popular.

Molecular weight sizing of proteins after blot development This product was previously called Prism Ultra Protein Ladder (10 - 180 kDa). 5 Steps to Multiplexed Fluorescent Western Blotting.

(B) lysate sonication is critical to the observation of nuclear and chromatin-bound proteins. It is useful to include both types of markers for both monitoring experiment progression and ease of interpreting results. Add Tween 20 detergent to the wash buffer to a final concentration of 0.05. Using a biotinylated marker offers visualization of the ladder along with antibody targets on film. Increase the number of washes and/or the volume of buffer used.

Monitoring blot transfer efficiency with easy orientation via multicolor standards Use Thermo Scientific SuperSignal Western Blot Enhancer to reduce background and enhance detection of low-abundance and weakly immunoreactive antigens.

These standards contain ten Strep-tagged recombinant proteins (10–250 kD), including three pink reference bands (25, 50, and 75 kD), as well as unstained versions of the Strep-tagged recombinant proteins for molecular weight sizing after blot development.

#Protein ladder for western blot plus

Additional data should be considered for a more accurate adjustment.Ġ.2~0.4 mg/ml of each band of protein is stored in a mixture of solution that contains: 20 mM Trisphosphate at pH 7.5, 2% SDS, 0.2 mM Dithiothreitol, 3.6 M Urea, and 15% (v/v) Glycerol.ĥμl of IRIS9 Prestained Protein Ladder (PM009) resolves 9 bands in 4-20% SDS-PAGE (Tris-glycine buffer) and after Western blotting to PVDF membrane.Use Precision Plus Protein WesternC Standards as a combination standard for western blotting and fluorescence detection in SDS-PAGE. Note: The molecular weight of each protein (kDa) was measured against an unstained protein ladder in every electrophoresis condition. Each protein can be stained with a different color for easy identification or labeled with affinity tags for western blot detection. 2.5 μl per well for general Western transferring. Recombinant protein ladders are engineered to produce tight bands, evenly spaced molecular weights, and other specific traits. 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. Refer to the IRIS9 Prestained Protein Ladder patterns in various electrophoresis conditions:ģ μl or 5 μl per loading for clear visualization during electrophoresis on 15-well or 10-well mini-gel, respectively. Western blot protein ladders are designed for protein molecular weight estimation directly on the blot during detection. Guide for Molecular Weight Estimation (kDa) The IRIS9 Prestained Protein Ladder keeps track of the size and separation of proteins during SDS-polyacrylamide gel electrophoresis, approximating the target protein size and validating the Western transfer efficiency on PVDF, nylon, or nitrocellulose membranes.

The 9 recombinant proteins are covalently coupled with blue chromophore, while 2 red bands at 70 kDa and 30 kDa, a green band at 15 kDa and a newly designed peacock green band at 60 kDa serve as reference bands. The IRIS9 Prestained Protein Ladder is a combination of 9 pre-stained proteins with molecular weights from 15 to 180 kDa.