Top 10 Most-read Protocols
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Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation
Authors:  Esteban A. Orellana and Andrea L. Kasinski, date: 11/05/2016, view: 9819, Q&A: 0
The SRB assay has been used since its development in 1990 (Skehan et al., 1990) to inexpensively conduct various screening assays to investigate cytotoxicity in cell based studies (Vichai and Kirtikara, 2006). This method relies on the property of SRB, which binds stoichiometrically to proteins under mild acidic conditions and then can be ...
Analysis of Protein Stability by the Cycloheximide Chase Assay
Comparison of protein stability in eukaryotic cells has been achieved by cycloheximide, which is an inhibitor of protein biosynthesis due to its prevention in translational elongation. It is broadly used in cell biology in terms of determining the half-life of a given protein and has gained much popularity in cancer research. Here we present a ...
BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry
Authors:  Bo Qiu and M. Celeste Simon, date: 09/05/2016, view: 8798, Q&A: 0
Lipid droplets (LDs) are ubiquitous, dynamic organelles and function as a storage depot for neutral lipids, including triglycerides and cholesterol esters (Walther and Farese, 2012). The movement of lipid species into and out of LDs impacts a variety of cellular processes, such as energy homeostasis, lipid-based signaling, and membrane homeostasis ...
Immunofluorescent Staining of Mouse Intestinal Stem Cells
Authors:  Kevin P. O’Rourke, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 9011, Q&A: 0
Immunofluorescent staining of organoids can be performed to visualize molecular markers of cell behavior. For example, cell proliferation marked by incorporation of nucleotide (EdU), or to observe markers of intestinal differentiation including paneth cells, goblet cells, or enterocytes (see Figure 1). In this protocol we detail a method to fix, ...
Sucrose Preference Test to Measure Stress-induced Anhedonia
Authors:  Andrew L. Eagle, Michelle Mazei-Robison and Alfred J. Robison, date: 06/05/2016, view: 7844, Q&A: 0
The 2-bottle choice procedure for assessing sucrose preference is a useful test to investigate anhedonia (i.e., inability to feel pleasure) in laboratory rodents, particularly in stress-based models of depression. The 2-bottle choice procedure allows for a comparison between behavioral preference for sucrose solution in drinking water ...
Differentiation of THP1 Cells into Macrophages for Transwell Co-culture Assay with Melanoma Cells
Authors:  Michael Peter Smith, Helen Young, Adam Hurlstone and Claudia Wellbrock, date: 11/05/2015, view: 11446, Q&A: 0
Understanding how immune cells such as macrophages interact with cancer cells is of increasing interest, as cancer treatments move towards combing both targeted- and immuno-therapies in new treatment regimes. This protocol is using THP-1 cells, a human leukemia monocytic cell line that can be differentiated into macrophages. This allows studying ...
Protocol-In vitro T Cell Proliferation and Treg Suppression Assay with Celltrace Violet
Authors:  Kristofor K. Ellestad and Colin C. Anderson, date: 01/05/2016, view: 7816, Q&A: 0
Measurement of the incorporation of radionuclides such as 3H-thymidine is a classical immunological technique for assaying T cell proliferation. However, such an approach has drawbacks beyond the inconvenience of working with radioactive materials, such as the inability of bulk radionuclide incorporation measurements to accurately ...
Plant Tissue Trypan Blue Staining During Phytopathogen Infection
In this protocol plant tissue is stained with trypan blue dye allowing the researcher to visualize cell death. Specifically this method avoids the use of the carcinogen compound chloral hydrate, making this classical method of staining safer and faster than ever. The protocol is applied specifically to detect cell death on Arabidopsis ...
Lung Section Staining and Microscopy
Authors:  Xiaofeng Zhou and Bethany B Moore, date: 05/20/2017, view: 3267, Q&A: 0
Our protocol describes immunofluorescent staining, hematoxylin and eosin staining and Masson’s trichrome staining on lung sections.
In vivo Bioluminescence Imaging of Luciferase-labeled Cancer Cells
Over the past decade, in vivo bioluminescent imaging has emerged as a non-invasive and sensitive tool for studying ongoing biological processes within living organisms (Contag et al., 1997; Contag et al., 1998). Based on the detection and quantitation of the photons produced by the oxidation of luciferin by luciferase ...