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AB139480

P4HB Inhibitor Screening Assay Kit

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(2 Publications)

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.

View Alternative Names

ERBA2L, PDI, PDIA1, PO4DB, P4HB, Protein disulfide-isomerase, Cellular thyroid hormone-binding protein, Prolyl 4-hydroxylase subunit beta, p55

3 Images
Functional Studies - P4HB Inhibitor Screening Assay Kit (AB139480)
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Supplier Data

Functional Studies - P4HB Inhibitor Screening Assay Kit (AB139480)

Intra-plate and inter-plate reproducibility using Inhibitor Control (bacitracin).

Dose response assay was performed with 0 to 3000 μM bacitracin added 15 minutes prior to the initiation of enzymatic reaction. Reactions were performed as described in Methods and Procedures section. Intra-plate and inter-plate CVs using the assay are typically 3-6%.

Functional Studies - P4HB Inhibitor Screening Assay Kit (AB139480)
  • FuncS

Supplier Data

Functional Studies - P4HB Inhibitor Screening Assay Kit (AB139480)

Assay validation using Inhibitor Control (bacitracin).

Dose response assay was performed with 0 to 3000 μM bacitracin added 15 min prior to the initiation of enzymatic reaction. Reactions were performed as described in Assay Protocol section. The fluorescence-based assay provides a vastly improved assay signal window and improved lower detection limit, In addition, the Z'-factor score obtained using the assay (0.91 for assay with and without P4HB) demonstrates excellent signal-to-noise and signal-to-background ratio

Functional Studies - P4HB Inhibitor Screening Assay Kit (AB139480)
  • FuncS

Supplier Data

Functional Studies - P4HB Inhibitor Screening Assay Kit (AB139480)

Absorption and fluorescence emission spectra for P4HB (PDIA1) Detection Reagent.

All spectra were determined in PBE Buffer. Example data only.

Key facts

Detection method

Fluorescent

Sample types

Purified protein, Inhibitor compounds

Assay type

Enzyme activity

Sensitivity

= 37 µM

Assay Platform

Microplate reader

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Enzyme activity assay": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

{ "values": { "2x96Tests": { "sellingSize": "2 x 96 Tests", "publicAssetCode":"ab139480-2x96Tests", "assetComponentDetails": [ { "size":"1 x 25 mL", "name":"PBE Buffer", "number":"AB139480-CMP04", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Inhibitor Control (Bacitracin)", "number":"AB139480-CMP02", "productcode":"" }, { "size":"1 x 5 mL", "name":"Deionized Water", "number":"AB139480-CMP01", "productcode":"" }, { "size":"2 x 165 µL", "name":"P4HB (PDIA1) (Human, Recombinant)", "number":"AB139480-CMP06", "productcode":"" }, { "size":"2 x 1.3 mL", "name":"DTT", "number":"AB139480-CMP08", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Insulin (from bovine pancreas)", "number":"AB139480-CMP03", "productcode":"" }, { "size":"1 x 1 mL", "name":"Stop Reagent", "number":"AB139480-CMP07", "productcode":"" }, { "size":"1 x 20 µL", "name":"P4HB (PDIA1) Detection Reagent", "number":"AB139480-CMP05", "productcode":"" } ] } } }
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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
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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.
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Product protocols

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Target data

This multifunctional protein catalyzes the formation, breakage and rearrangement of disulfide bonds. At the cell surface, seems to act as a reductase that cleaves disulfide bonds of proteins attached to the cell. May therefore cause structural modifications of exofacial proteins. Inside the cell, seems to form/rearrange disulfide bonds of nascent proteins. At high concentrations and following phosphorylation by FAM20C, functions as a chaperone that inhibits aggregation of misfolded proteins (PubMed : 32149426). At low concentrations, facilitates aggregation (anti-chaperone activity). May be involved with other chaperones in the structural modification of the TG precursor in hormone biogenesis. Also acts as a structural subunit of various enzymes such as prolyl 4-hydroxylase and microsomal triacylglycerol transfer protein MTTP. Receptor for LGALS9; the interaction retains P4HB at the cell surface of Th2 T helper cells, increasing disulfide reductase activity at the plasma membrane, altering the plasma membrane redox state and enhancing cell migration (PubMed : 21670307).
See full target information P4HB
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Publications (2)

Recent publications for all applications. Explore the full list and refine your search

Scientific reports 9:18300 PubMed31797966

2019

Male-Specific Protein Disulphide Isomerase Function is Essential for Plasmodium Transmission and a Vulnerable Target for Intervention.

Applications

Unspecified application

Species

Unspecified reactive species

Fiona Angrisano,Katarzyna A Sala,Sofia Tapanelli,George K Christophides,Andrew M Blagborough

Frontiers in chemistry 2:73 PubMed25232538

2014

Methods of measuring protein disulfide isomerase activity: a critical overview.

Applications

Unspecified application

Species

Unspecified reactive species

Monica M Watanabe,Francisco R M Laurindo,Denise C Fernandes
View all publications
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websiteProtocolBooklet
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