This allows you to easily distinguish bands representing your protein of interest from those representing your housekeeping protein.Įnsure your loading control is highly expressed in your sample and its level remains unchanged throughout an experiment. Don’t use a housekeeping gene that you’ve found to vary in expression level during your experimental treatment previously. Probing with a loading control can be carried out alongside the target antibody by cutting the membrane before antibody incubation, or after the blot has been stripped of previously bound antibodies. Perform titration experiments on your samples to determine the concentration of antibody against your housekeeping protein and blot exposure time that gives optimum signal-to-background ratio without signal saturation. Housekeeping proteins are often highly expressed, making it easy for the signal from the housekeeping protein detection to become saturated and affect normalization. If you are performing a multiplexed fluorescent western blot experiment, choose your brightest fluorophore for your protein of interest and your dimmest fluorophore for your housekeeping protein. This will prevent overexposure and therefore oversaturation of the signal from your housekeeping protein.īeta Actin is a 42 kDa protein ubiquitously expressed in all eukaryotic cells. "Image Studio Lite Quantification Software has been discontinued.It plays a critical role in the regulation of cell migration and gene expression through control of the cellular G-actin pool (Bunnell et al."Quantification of Western Blots Using ImageJ"."Analyzing gels and western blots with ImageJ". "Quantifying western blots without expensive commercial quantification software". "Reciprocal Interferences of TNF-α and Wnt1/β-Catenin Signaling Axes Shift Bone Marrow-Derived Stem Cells Towards Osteoblast Lineage after Ethanol Exposure". "Type I Interferon Protects against Pneumococcal Invasive Disease by Inhibiting Bacterial Transmigration across the Lung". "A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression". "ER Stress Activating ATF4/CHOP-TNF-a Signaling Pathway Contributes to Alcohol-Induced Disruption of Osteogenic Lineage of Multipotential Mesenchymal Stem Cell". "Chronic exposure to benzyl butyl phthalate (BBP) alters social interaction and fear conditioning in male adult rats: Alterations in amygdalar MeCP2, ERK1/2 and ERα". Betz, A Jayatilaka, S Joshi, J Ramanan, S Debartolo, D Pylypiw, H Franke, E (2013)."Suppression of type 1 pilus assembly in uropathogenic Escherichia coli by chemical inhibition of subunit polymerization". Journal of the American Heart Association. "Functional, Anatomical, and Molecular Investigation of the Cardiac Conduction System and Arrhythmogenic Atrioventricular Ring Tissue in the Rat Heart". "Densitometry Tutorial for Image Studio Lite from LI-COR"."Western Blot Quantification with LI-COR Image Studio Lite"."Analyzing western blots with Image Studio Lite". "Image Studio Lite: Tutorial Guide" (PDF).Release history of Image Studio Lite software: The primary function of Image Studio Lite is to quantitate relative abundance of proteins on a Western blot or DNA/RNA in an electrophoresis gel from an image of the blot or gel. It is unclear from this announcement whether Empiria Studio will support acquisition for Odyssey FC imaging systems previously using Image Studio or if licenses will be granted to owners of machines previously using Image Studio. In its place, LICOR has released Empiria Studio, a commercial product. Image Studio™ Lite is free image processing software used for quantitation of Western blot images and images from related experiments.Īs of 2021, Image Studio Lite has been discontinued and is no longer available for download. bio /products /software /image _studio _lite /
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