Generation and Testing of Fluorescent Adaptable Simple Theranostic (FAST) Proteins
[Abstract] This protocol provides a step-by-step method to create recombinant fluorescent fusion proteins that can be secreted from mammalian cell lines. This builds on many other recombinant protein and fluorescent protein techniques, but is among the first to harness fluorescent fusion proteins secreted directly into cell culture supernatant. This opens ...
Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)
[Abstract] The binding interactions of PD-1 and PD-L1 have been studied by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but these investigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the ...
In situ, Cell-free Protein Expression on Microarrays and Their Use for the Detection of Immune Responses
Authors: Katrin Hufnagel,
Dennis Reininger,
Siu Wang Ng,
Nadine Gassert,
Juliane K. Rohland,
Soroosh Shahryarhesami,
Andrea S. Bauer,
Tim Waterboer and
Jörg D. Hoheisel,
date: 02/05/2019,
view: 2872,
Q&A: 0
[Abstract] Until recently, whole-proteome microarrays for comprehensive studies of protein interactions were mostly produced by individual cloning and cellular expression of very many open reading frames, followed by protein isolation and purification as well as array production. To overcome this cumbersome process, we have developed a method to generate ...
Adoptive Transfer of Monocytes Sorted from Bone Marrow
[Abstract] Inflammatory Ly6Chi monocytes can give rise to distinct mononuclear myeloid cells in the tumor microenvironment, such as monocytic myeloid-derived suppressor cells (Mo-MDSC), immature macrophages, M2-like tumor-associated macrophages (TAMs), M1-like TAMs or monocyte-derived dendritic cells (Mo-DCs). This protocol describes a method to ...
In-vivo gp100-specific Cytotoxic CD8+ T Cell Killing Assay
[Abstract] Cytotoxic CD8+ T lymphocytes (CTLs) represent a crucial component of the adaptive immune system and play a prominent role in the anti-tumor immune responses of both mice and humans.
Cytotoxic CD8+ T cells are responsible for the lysis of cells expressing peptides associated with MHC class I molecules and derived ...
Glioma Induction by Intracerebral Retrovirus Injection
Authors: Ravinder K Verma ,
Fanghui Lu and
Qing Richard Lu,
date: 07/20/2017,
view: 6183,
Q&A: 0
[Abstract] Glioblastoma (GBM) is the most common primary brain cancer in adults and has a poor prognosis. It is characterized by a high degree of cellular infiltration that leads to tumor recurrence, atypical hyperplasia, necrosis, and angiogenesis. Despite aggressive treatment modalities, current therapies are ineffective for GBM. Mouse GBM models not only ...
In vivo Efficacy Studies in Cell Line and Patient-derived Xenograft Mouse Models
[Abstract] In vivo xenograft models derived from human cancer cells have been a gold standard for evaluating the genetic drivers of cancer and are valuable preclinical models for evaluating the efficacy of cancer therapeutics. Recently, patient-derived tumorgrafts from multiple tumor types have been developed and shown to more accurately ...
Quantification of Tumor Material Uptake
Authors: Richard N. Hanna and
Catherine C. Hedrick,
date: 10/20/2016,
view: 6232,
Q&A: 0
[Abstract] Extracellular tumor material including exosomes, microvesicles and apoptotic tumor debris may help cancers invade new organs. Enhancing the removal of extracellular tumor material by immune cells represents a novel immunotherapy approach for preventing cancer metastasis. This protocol quantifies the uptake and removal of extracellular tumor ...