Search
Top 10 Most-read Protocols
Most Read rankings are calculated based on published articles (since 2015) read in the last two months. Rankings are calculated monthly. Rankings are based on page view data provided by Google Analytics.
In vitro Assays for the Detection of Calreticulin Exposure, ATP and HMGB1 Release upon Cell Death
Authors:  Yuting Ma and Heng Yang, date: 12/20/2016, view: 2753, Q&A: 0
Accumulating evidence is revealing the essential role of immune system in cancer treatment. Certain chemotherapeutic drugs can potently induce the release of ‘cell death associated molecular patterns’ (CDAMPs), which accompanies cancer cell demise. CDAMPs can engage corresponding receptors on immune cells and stimulate immune responses to achieve ...
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 ...
Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation
Authors:  Esteban A. Orellana and Andrea L. Kasinski, date: 11/05/2016, view: 7757, 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 ...
Skin Wound Healing Model - Excisional Wounding and Assessment of Lesion Area
This protocol focus on the most common surgical mouse model of cutaneous excisional wound healing used to study the cellular and molecular pathways involved in wound repair and regeneration as well as in translational applications such as the evaluation of new therapeutic modalities. This model allows the monitoring of the wound closure and the ...
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: 7334, 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 ...
Sucrose Preference Test to Measure Stress-induced Anhedonia
Authors:  Andrew L. Eagle, Michelle Mazei-Robison and Alfred J. Robison, date: 06/05/2016, view: 6693, 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 ...
Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria
Authors:  Tomáš Zavřel, Maria A. Sinetova and Jan Červený, date: 05/05/2015, view: 8500, Q&A: 0
This is a protocol for precise measurement of chlorophyll a and total carotenoid concentrations in cyanobacteria cells. Cellular chlorophyll concentration is one of the central physiological parameters, routinely followed in many research areas ranging from stress physiology to biotechnology. Carotenoids concentration is often related to ...
Immunofluorescent Staining of Mouse Intestinal Stem Cells
Authors:  Kevin P. O’Rourke, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 7497, 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, ...
Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells
Authors:  Kevin P. O’Rourke, Sarah Ackerman, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 6902, Q&A: 3
In this protocol we describe our modifications to a method to isolate, culture and maintain mouse intestinal stem cells as crypt-villus forming organoids. These cells, isolated either from the small or large intestine, maintain self-renewal and multilineage differentiation potential over time. This provides investigators a tool to culture wild ...
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: 6620, 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 ...