Tumor Lysate Preparation
Tumor Lysate Preparation - Related Videos
03:36
Preparation of Bacteriophage Lysate for Generalized Transduction
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2026
Source: Saragliadis, A., et al. Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes. J. Vis. Exp.(2018)This video demonstrates preparation of a bacteriophage lysate containing transducing particles. A donor strain is infected, plaques form, and the soft agar overlay is processed with chloroform to yield a clarified lysate for downstream analysis.
03:17
Preparing Organoids from Tumors: A 3D In Vitro Tumor Model
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2023
Organoids derived from tumors are a type of 3D cell culture model that can maintain the pathological features and organ-specific cell lineage of tumors. In the example protocol, we will see scientists generate organoids from a prostate tumor from a genetically engineered mouse model.
04:08
Purification of Bacteriophages from Bacterial Lysate
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2026
Source: Pett, N., et al. T4 Bacteriophage and E. coli Interaction in the Murine Intestine: A Prototypical Model for Studying Host-Bacteriophage Dynamics In Vivo. J. Vis. Exp. (2024)This video demonstrates the purification of bacteriophages from bacterial cell lysates using sequential centrifugation, membrane filtration, chloroform extraction, centrifugal filtration, and buffer exchange to obtain a purified phage preparation.
08:32
Preparation of Pooled Human Platelet Lysate (pHPL) as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures
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Cited by 133 •
2009
Human platelet lysate is a rich source of growth factors and a potent supplement in cell culture. This protocol presents the process of preparing a large pool of human platelet lysate by starting from platelet rich plasma, performing several freeze-thaw cycles and depleting the platelet fragments.
08:33
Cell-Free In Vitro mRNA Translation In Drosophila Lysates
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2026
Here, we provide details on how to prepare lysates and perform in vitro translation using Drosophila embryos.
FAQs
Controlled, cold processing helps preserve soluble proteins and other antigenic components that could be altered during uncontrolled tissue disruption. Mechanical or physical lysis must therefore release cellular material without unnecessarily compromising the signals recognized by immune cells. Maintaining these conditions improves the preparation’s suitability for downstream antigen presentation, dendritic-cell stimulation, and immune-response analyses.
Clarification separates insoluble debris from the soluble fraction containing retained antigenic material. This produces a more consistent extract for immunological assays and reduces particulate material that could interfere with handling or analysis. The clarified preparation can then be quantified and divided into aliquots, supporting more reproducible comparisons between experiments or immune-response measurements.
A broad antigen mixture allows investigators to examine immune recognition of complex tumor material rather than restricting analysis to one selected antigen. This is useful when the goal is to evaluate overall antigen presentation or responses involving antibodies and T cells. The same feature supports assessment of how immune cells respond to diverse components from tumor tissue or cultured tumor cells.
The workflow begins with tumor tissue or cultured tumor cells, followed by controlled mechanical or physical disruption under cold conditions. The disrupted material is clarified to remove insoluble debris, after which the resulting lysate is quantified, aliquoted, and stored for later use. These stages create a prepared extract that can be incorporated consistently into downstream immune assays.
Quantification helps investigators characterize the amount of prepared material available for an experiment, while aliquoting supports consistent handling across repeated assays. Storage makes the preparation available for later analysis without repeatedly disrupting new tumor material. Together, these steps facilitate experimental organization and comparison of antigen presentation, dendritic-cell stimulation, antibody, or T-cell response studies.
Tumor lysates can be incorporated into antigen presentation assays and dendritic-cell stimulation experiments to examine how immune cells process or respond to complex tumor material. They also support analysis of antibody and T-cell responses and evaluation of tumor vaccines. These applications help investigators characterize immune recognition and compare responses to broadly represented tumor-associated antigens.