Ptm Profiling
Ptm Profiling - Related Videos
06:26
PTM Based Tumor Microenvironment Monitoring: A Method for Tracking Changes in ECM Rigidity Using Tumor Spheroids
0 Views •
2023
This protocol describes a technique for monitoring extracellular matrix rigidity changes by using particle-tracking microrheology (PTM). This is done by culturing 3D pancreatic tumor spheroids into ECM with embedded fluorescent probes for video particle-tracking and analysis.
10:54
Extraction of Histones from Clinical Specimens for Epigenetic Profiling by Mass Spectrometry
0 Views •
2025
This protocol enables efficient histone extraction from various clinical samples for subsequent post-translational modification analysis by LC-MS/MS. By facilitating robust detection of key histone modifications, it serves as a valuable tool for comprehensive epigenetic profiling in large clinical cohorts.
05:21
A Peptide Microarray Technology for Specificity Profiling of Antibodies
0 Views •
2025
This video demonstrates a technique that applies peptide microarray technology to profile the specificity of antibodies recognizing post-translational modifications (PTMs) on histone peptides. A microarray slide containing immobilized fluorophore-conjugated histone peptides with known PTM combinations is treated with target PTM-specific antibodies. This process helps identify the antibody's specificity for the target and its potential cross-reactivity with non-target PTMs.
08:29
Expression Profiling with Microarrays
0 Views •
2023
Microarrays are important tools for profiling gene expression, and are based on complementary binding between probes that are attached to glass chips and nucleic acids derived from samples. Using these arrays, scientists can simultaneously evaluate the expression of thousands of genes. In addition, the expression profiles of different cells or tissue types can be compared, allowing researchers to deduce how the expression of different genes change during biological processes, and thus gain...
07:13
Using Topographic Maps to Generate Topographic Profiles
0 Views •
2023
Source: Laboratory of Alan Lester - University of Colorado Boulder Topographic maps are "plan-view" representations of Earth's three-dimensional surface. They are a standard type of map-view that provides an overhead, or aerial, perspective. Among the defining features of a topographic map are the contour lines that indicate locations of constant elevation. The elevation interval between the contour lines is dependent on the level of detail provided by the map and the kind of topography present.
FAQs
Enrichment focuses analysis on modified species before instrument measurement. By concentrating modified peptides or proteins, the workflow directs subsequent liquid-chromatography separation and mass-spectrometric analysis toward relevant targets within a biological sample. This organization supports more effective detection of modification sites and relative abundances associated with regulatory changes.
Mass and fragmentation patterns provide complementary evidence for assigning a modification and its location. Mass spectrometry first measures the analyzed species, while fragmentation generates patterns that help identify the modified peptide region. Together, these measurements allow researchers to move beyond detecting a modification and determine which site on the protein carries it.
Relative abundance indicates how strongly a particular modified species is represented in one sample compared with another or within the measured set. Changes in these values can reveal altered regulatory events across cellular states or after experimental treatment. Interpreting abundance alongside modification identity and site assignment helps connect molecular changes with biochemical responses.
Examining several modification classes broadens the view of protein regulation because these chemical changes can be associated with effects on activity, stability, localization, and interactions. A combined profile can therefore reveal multiple regulatory events within the same biological context, helping biochemists relate coordinated changes in protein modification to signaling pathways or cellular states.
A typical workflow begins with a biological sample, followed by enrichment of modified peptides or proteins when appropriate. The enriched material is separated by liquid chromatography, then introduced to mass spectrometry for mass measurement and fragmentation. The resulting data are used to assign modification types, sites, and relative abundances across the samples examined.
The workflow centers on biological samples containing modified proteins, peptides or proteins selected through enrichment, liquid-chromatography separation, and a mass spectrometer capable of measuring mass and fragmentation patterns. These components work in sequence: enrichment focuses the material, chromatography separates it, and mass spectrometry supplies the evidence needed for modification and site assignment.
Researchers use this approach when they need to connect protein regulation with a cellular state, disease mechanism, or response to an experimental treatment. The measurements can identify changes in phosphorylation, acetylation, ubiquitination, and other modifications, then relate those events to signaling pathways and protein function. This makes profiling useful for comparing biologically different sample conditions.