Description
Key facts
Detection method:Fluorescent,
Sample types:Purified protein, Inhibitor compounds,
Assay type:Enzyme activity,
Sensitivity:= 37 µM,
Assay Platform:Microplate reader
Product details:
Abcam's P4HB (PDIA1) Inhibitor Screening Assay Kit (ab139480) provides a simple, homogenous assay for screening modulators of protein disulfide isomerase (PDIA1) enzymatic activity in a 96-well microplate. The basis for the assay is the P4HB (PDIA1)-catalyzed reduction of insulin in the presence of DTT, resulting in the formation of insulin aggregates which in turnn bind to the red-emitting fluorogenic P4HB (PDIA1) Detection Reagent.
Relative to analogous turbidimetric assays of P4HB (PDIA1) activity, the fluorescence-based assay provides a vastly improved assay signal window, improved lower detection limit and superior Z'-score (>0.8).
The kit includes human recombinant P4HB (PDIA1) enzyme as positive control, as well as the P4HB (PDIA1) inhibitor bacitracin as a negative control.
This kit will provide a quantitative readout of P4HB (PDIA1) enzymatic activity is a robust and HTP fashion and can be applied to identification of P4HB (PDIA1) inhibitors from chemical libraries.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions--80°C, Appropriate long-term storage conditions--80°C, Storage information--80°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
P4HB is a protein known as Protein Disulfide Isomerase (PDI). It plays a significant role in catalyzing the formation of disulfide bonds. This protein helps to fold newly synthesized proteins by shuffling disulfide bonds. It has a molecular mass of around 57 kDa. P4HB is mainly expressed in the endoplasmic reticulum of cells. While its alternative name is ERp59 P4HB is integral to the protein folding machinery inside cells.
Biological function summary
P4HB functions in protein folding and assembly. By acting as a chaperone it protects proteins from misfolding and aggregation. P4HB operates as part of a larger multi-protein complex that assists in maintaining protein structure under stress conditions in the cell. Its activity ensures protein stability and proper cellular function important for cell viability and health.
Pathways
P4HB plays a central role in the unfolded protein response (UPR) and oxidative protein folding pathway. It interacts with proteins such as calnexin and calreticulin through its involvement in these pathways. During oxidative protein folding P4HB introduces disulfide bonds into nascent proteins while removing incorrect ones ensuring efficient protein quality control.
P4HB alteration has connections to diseases like cancer and neurodegenerative disorders. Its overexpression and activity can promote tumor growth by aiding cancerous cell survival through protein homeostasis. Moreover P4HB's association with amyloid precursor protein impacts the progression of Alzheimer's disease. These connections make P4HB a relevant target for research into therapeutic interventions.
