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Isothermal Titration Calorimetry: A Biophysical Method to Characterize the Interaction between Label-free Biomolecules in Solution

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A Lentiviral Pseudotype ELLA for the Measurement of Antibodies Against Influenza Neuraminidase

Authors: Fabrizio Biuso, George Carnell, Emanuele Montomoli and Nigel Temperton, date: 07/20/2018, view: 230, Q&A: 0

This protocol describes the rapid and safe production of lentiviral pseudotypes characterized by a lentiviral core containing a reporter, in conjunction with avian influenza haemagglutinin (HA) and human neuraminidase (NA) glycoproteins on the ...

In vitro Chaperone Activity Assay Using α-Amylase as Target Protein

Authors: Jeeshma Nambidi Parambath, Gayathri Valsala, Karthik Menon and Shiburaj Sugathan, date: 06/20/2018, view: 378, Q&A: 0

Small heat shock proteins (sHSP) are stress proteins which are ubiquitously found in almost all living organisms. They function as molecular chaperones, which assist in protein folding during translation and in the prevention of irreversible protein ...

Purifying Properly Folded Cysteine-rich, Zinc Finger Containing Recombinant Proteins for Structural Drug Targeting Studies: the CH1 Domain of p300 as a Case Example

Authors: Yong Joon Kim, Stefan Kaluz, Anil Mehta, Emily Weinert, Shannon Rivera and Erwin G. Van Meir, date: 09/05/2017, view: 2437, Q&A: 0

The transcription factor Hypoxia-Inducible Factor (HIF) complexes with the coactivator p300, activating the hypoxia response pathway and allowing tumors to grow. The CH1 and CAD domains of each respective protein form the interface between p300 and ...

Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-histone Antibodies

Author: Zheng Liu, date: 06/20/2012, view: 7360, Q&A: 0

Autoimmune disease is characterized with the break of tolerance against self antigens. Anti-histone antibodies can be found in the majority of patients with system lupus erythematosus (SLE) and a number of lupus-prone mouse strains. This protocol ...

RNP-IP (Original Method): Obtaining Majority RNA from RNA Binding Protein in the Nucleus

Author: FengZhi Liu, date: 05/20/2012, view: 14322, Q&A: 1

Post-transcriptional regulation of gene expression is a ribonucleoprotein (RNP)-driven process, which involves RNA binding proteins (RBPs) and noncoding RNAs that regulate splicing, nuclear export, subcellular localization, mRNA stability and ...

Antibody Affinity Purification

Author: Huan Pang, date: 05/05/2012, view: 8816, Q&A: 0

Antibody purification is performed to concentrate and enrich antigen-specific antibodies and lower the background signal during detection by removing any non-specific proteins. Affinity purification makes use of specific binding interactions between ...

Immunofluorescence Detection and F-actin Staining of MTLn3 Cells

Author: Huan Pang, date: 04/20/2012, view: 10425, Q&A: 0

Epidermal growth factor (EGF)-stimulated MTLn3 cells protrusion play an important role in cell migration. Phalloidin which binds F-actin in cells is an imaging tool used with light microscopy to investigate the distribution of actin. The protocol ...

PI (phosphoinositide)-3 Kinase Assay

Author: Huan Pang, date: 04/20/2012, view: 7192, Q&A: 1

PI3-kinases regulate a wide range of cellular responses through the production of phosphatidylinositol 3, 4, 5-trisphosphate [PI (3, 4, 5) P (3)] in cellular membranes. This protocol provides a highly efficient and easy-to-handle method for ...

Cell Cycle Analysis Using Propidium Iodide Staining with GFP Detection

Author: Hui Zhu, date: 04/05/2012, view: 16005, Q&A: 3

Infecting mammalian cells with a GFP construct to overexpress or knockdown target genes is one of the most commonly used methods to study and manipulate gene expression. To determine the target gene function on the cell cycle, analyzing the cell ...

Protein Extraction from Mice Xenograft Tumor

Author: Huan Pang, date: 04/05/2012, view: 14669, Q&A: 1

Proteins from frozen tissue such as xenograft tumor need to be extracted efficiently without being degraded to make the best use of a limited resource and to ensure that an accurate representation of proteins in selected tumor tissue can be ...