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Western Blot Services
Western blots remain the foremost biochemical method for protein analysis. The specificity of the Western blot protocol makes it ideal for protein quantitation.
Our Western blot services provide robust and flexible options for detecting your protein of interest in a variety of sample types. Customized Western blot workflows can accommodate your experimental needs.
Variable Gel Chemistry
Gel chemistry can be modified to suit your needs. Native, denaturing, and gradient gels are easily incorporated into our Western blot services.
Antibody Options Available
Our existing collection of vetted primary and secondary antibodies are available for use. We gladly utilize client-supplied reagents as well.
Co-Immunoprecipitation Studies
Probe protein interactions with expertly executed co-immunoprecipitation (co-IP) studies. Co-IPs can characterize known interacting partners, or identify novel interacting proteins.
Western Blots of Subcellular Fractions
To determine intracellular localization, cell lysates can be fractionated and analyzed as part of your Western blot workflow.
Sample Flexibility
Custom Western blot procedures can be devised to process complex biological samples or incompatible sample reagents prior to immunoblotting.
Comprehensive Project Support
Ichor provides comprehensive Western blot services to ensure that your project receives full technical and scientific support throughout experimental execution.
Complete Protein Analysis Capabilities
Ichor’s Western blot services are not limited to a standard protocol. We provide many variations as part of our full-service biochemical assay offerings. These include total protein analyses, dot blots, native gel chemistry, zymograms, interaction studies, protein band isolation, recombinant protein production and analysis, and more. Ichor can integrate these analyses into a larger experimental workflow to optimize project workflow and reduce experimental timelines.
Achieve High-Quality Western Blot Results
Featured Case Studies / Capabilities
Quantitative Western Blot Analysis
Protein Half-Life Determination
Western blotting can be used to quantitate a protein of interest from a complex sample. Absolute quantitation requires a standard curve of known protein amounts, while relative quantitation can be performed on any Western blot by densitometry calculation. Relative quantitation can reveal trends in protein expression, modification, and stability.
Here, a modified Western blot procedure was used to determine the half-life of a recombinant protein in mammalian cell culture. Cells were transiently transfected with a construct for transgene expression, and cells were harvest over time for characterization of protein abundance. Densitometry was performed, providing relative quantitation at each time point. A one-phase decay model was applied to the data to determine protein half-life.
Better Phosphoprotein Detection
Cleaner Signaling Results
Characterizing protein phosphorylation provides critical insight into intracellular signaling cascades. Western blot analysis is a gold standard method for quantitating phosphoprotein abundance, but also comes with inherent drawbacks. Polyclonal antibodies for phosphoproteins often bind nonspecifically to other intracellular proteins, creating background and detracting from identification and quantitation of the antigen of interest.
Our scientists are experts in phosphoprotein Western blot workflows. Through experience with difficult antibody-antigen pairs, we have devised protocol modifications to achieve clean phosphoprotein blots. By optimizing gel composition and blotting procedures, we maximize resolution of proteins of interest from non-specific bands, and minimize the appearance of background.
Western Blot Analysis of Membrane Proteins
Integral Membrane Protein Analysis
Western blot analysis for membrane proteins can often be difficult; the hydrophobicity of integral membrane proteins leads to poor solubilization from the membrane, or sample aggregation after isolation. Our experience has led to the development of several Western blot protocol modifications to promote successful resolution of membrane proteins. By altering gel and buffer composition, and incorporating additional solubilization steps, we can tackle difficult hydrophobic proteins.
Here, an integral membrane protein was overexpressed recombinantly in mammalian cells. Western blot analysis shows appropriate protein migration and good resolution, indicating successful protein solubilization.
Frequently Asked Questions
Read more about our Western blot services and capabilities.