Recombinant Human P4HB Protein, CF

R&D Systems | Catalog # 4236-DI

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human P4HB Protein (4236-DI)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Accession Number

P07237

Applications

Enzyme Activity
View all Protein Disulfide Isomerase/P4HB Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Protein Disulfide Isomerase/P4HB protein
Asp18-Lys505, with a C-terminal 10-His tag

Purity

>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Asp18

Predicted Molecular Mass

56 kDa

SDS-PAGE

60 kDa, reducing conditions

Activity

Measured by its ability to promote aggregation of insulin in the presence of DTT.
The specific activity is >7.5 A650/cm/min/mg, as measured under the described conditions.

Formulation, Preparation, and Storage

4236-DI
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Protein Disulfide Isomerase/P4HB

Protein Disulfide Isomerase, also known as prolyl 4‑hydroxylase subunit beta (P4HB), protocollagen hydroxylase, cellular thyroid hormone binding protein p55 and glutathione-insulin transhydrogenase (1-3) is an abundant multifunctional enzyme that belongs to the Protein Disulfide Isomerase family. It contains two thioredoxin domains that catalyze the formation, breakage and rearrangement of disulfide bonds. When present as a tetramer consisting of two alpha subunits and two beta subunits, this enzyme functions as a hydroxylase catalyzing the hydroxylation of prolyl residues in preprocollagen. P4HB has various additional functions (4-7). It binds thyroid hormone. It acts as a chaperone that inhibits aggregation of misfolded proteins. It plays a role in both the influx and efflux of S-nitrosothiol-bound nitric oxide. It is also a subunit of the microsomal triglyceride transfer protein complex.

References

  1. Pihlajaniemi, T. et al. (1987) EMBO J. 6:643.
  2. Cheng, S.Y. et al. (1987) J. Biol. Chem. 262:11221.
  3. Morris, J. I. and Varandani, P. T. (1988) Biochim. Biophys. Acta 949:169.
  4. Obata, T. et al. (1988) J. Biol. Chem. 263:782.
  5. Gilbert, H. F. (1997) J. Biol. Chem. 272:29399.
  6. Sliskovic, I. et al. (2005) J. Biol. Chem. 280:8733.
  7. Wetterau, J. R. et al. (1990) J. Biol. Chem. 265:9800.

Long Name

Proline 4-Hydroxylase, Beta Polypeptide

Alternate Names

DSI, ERBA2L, GIT, PDI, PDIA1, PO4DB, PO4HB, PROHB, Thbp

Entrez Gene IDs

5034 (Human); 18453 (Mouse); 25506 (Rat)

Gene Symbol

P4HB

UniProt

P07237

Additional Protein Disulfide Isomerase/P4HB Products

Product Documents for Recombinant Human P4HB Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human P4HB Protein, CF

For research use only

Related Research Areas

Redox Enzymes

Citations for Recombinant Human P4HB Protein, CF

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Protocols

View specific protocols for Recombinant Human P4HB Protein, CF (4236-DI):

Materials
  • Buffer A: 100 mM NaH2PO4, 3 mM DTT, pH 7.0
  • Buffer B: 100 mM NaH2PO4, 3 mM EDTA, pH 7.0
  • Recombinant Human Protein Disulfide Isomerase/P4HB (rhP4HB) (Catalog # 4236-DI)
  • Insulin (Sigma, Catalog # I-5500), 10 mg/mL solution in 50 mM Tris, pH 7.5 (solution will be cloudy)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute insulin to 1.5 mg/mL in Buffer B.
  2. Incubate diluted insulin for 10 minutes at room temperature (insulin will solubilize).
  3. Dilute rhP4HB to 45 µg/mL in Buffer A.
  4. Mix 100 µL of 45 µg/mL rhP4HB and 200 µL of 1.5 mg/mL insulin. Create a Substrate Blank with 100 µL Buffer A and 200 µL insulin.
  5. Incubate at room temperature for 20 minutes.
  6. Load in a clear 96-well plate 100 µL from each reaction vial in duplicate.
  7. Read at 650 nm (absorbance) in kinetic mode for 5 minutes.
  8. Calculate specific activity:

     Specific Activity (Abs/cm/min/mg) =

 Adjusted Vmax* (Abs/min)**
path corr.(cm)*** x amount of enzyme (mg)

     *Adjusted for Substrate Blank
     **Note: the output of many spectrophotometers in kinetic mode is in mOD.
     ***Using the pathlength correction of 0.32 cm

Per Well:
  • rhP4HB: 0.0015 mg
  • Insulin: 0.100 mg

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