Recombinant Human DPEP1 His-tag Protein, CF

R&D Systems | Catalog # 11524-DP

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human DPEP1 His-tag Protein (11524-DP)
  • 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

P16444.3

Applications

Enzyme Activity
View all DPEP1 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human DPEP1 protein
Asp17-Ser384, with C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Asp17

Predicted Molecular Mass

42 kDa

SDS-PAGE

39-49 kDa, under reducing conditions

Activity

Measured by its ability to cleave carnosine ( beta -Ala-L-His) in a two-step assay.
The specific activity is >5 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human DPEP1 His-tag Protein, CF

Recombinant Human DPEP1 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human DPEP1 His-tag Protein (Catalog # 11524-DP) was resolved with SDS-PAGE under reducing (R) condition and visualized by Coomassie® Blue staining, showing bands at 39-49 kDa, under reducing conditions.

Formulation, Preparation, and Storage

11524-DP
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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: DPEP1

Dipeptidase 1 (DPEP1), also referred to as renal peptidase or beta-lactamase, is a zinc-dependent, glycosylated member of the M19 peptidase metallo-dependent hydrolase superfamily that lacks a sequence commonly found in zinc proteases and has poor homology to other metallopeptiases (1). DPEP1 is a homodimeric elongated ellipsoid with a single disulfide bond between the subunits (1) and GPI-anchored to the cell membrane at the brush border of epithelial cells (2). Each DPEP1 monomer contains a zinc-dependent active site, a propeptide, and a GPI anchor at the c-terminus (1). DPEP1 has activity towards several dipeptides and is critical in glutathione and leukotriene metabolism as well as mammalian metabolism of the B-lactam ring of antibiotics (1, 3). DPEP1 expression is upregulated or associated as a marker in various cancers including colon, kidney, and prostate cancer (4-6). It is also upregulated in renal tubular epithelial cells and associated with iron-related ferroptotic cell death in diabetic retinopathy (2, 7) where it may mediate proinflammatory cancer mediatory leukotrienes or serve as a receptor as an adhesion receptor for neutrophil recruitment and impact inflammation (8-10). DPEP1 has been noted as a potential therapeutic marker or target for mestastasis, cancer progression, and disease in the liver and lungs with acute or chronic inflammation (2, 4, 9, 11).

References

  1. Nitanai, Y. et. al. (2002) J. Mol. Biol. 321:177.
  2. Li, Y. et. al. (2024) Int. Immunopharmacol. 133:111955. 
  3. Campbell, B.J. et. al. (1988) Biochim. Biophys. Acta 956:110.
  4. Tachibana, K. et. al. (2017) Biomed. Rep. 6:423.
  5. Xiang, C. et. al. (2023) J. Cell Mol. Med. 27:1947.
  6. Zhu, W. et. al. (2023) Heliyon 9:e18870.
  7. Allison, S.J. (2021) Nat. Rev. Nephrol. 17:707.
  8. Satoh, S. et. al. (1993) Biochim. Biophys. Acta 1172:181. 
  9. Choudhury, S.R. et. al. (2019) Cell 178:1205.
  10. Wang, M. (2022) Nat. Rev. Nephrol. 18:199. 
  11. Tutanov. O.S. et. al. (2023) Extracell. Vesicles Circ. Nucl. Acids. 4:195.

Long Name

Dipeptidase 1

Alternate Names

HRDP, Microsomal Dipeptidase, Renal Dipeptidase

Entrez Gene IDs

1800 (Human)

Gene Symbol

DPEP1

UniProt

P16444.3

Additional DPEP1 Products

Product Documents for Recombinant Human DPEP1 His-tag Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human DPEP1 His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human DPEP1 His-tag Protein, CF (11524-DP):

Materials
  • Assay Buffer: 50 mM Tris, pH 7.5
  • Recombinant Human DPEP1 (rhDEP1) (Catalog # 11524-DP)
  • Substrate: L-Carnosine, 100 mM stock in deionized water
  • Trichloroacetic acid (TCA), 10% stock in deionized water
  • o-Phthaldialdehyde (OPA), 50 mg/mL stock in DMSO
  • Sodium Hydroxide (NaOH), 2 M stock in deionized water
  • Black 96-well plate
  • Plate Reader with Fluorescence Read Capability
  1. Dilute rhDPEP1 to 20 µg/mL in Assay Buffer.
  2. Dilute Substrate to 4 mM in Assay Buffer.
  3. Dilute 10% TCA to 1% in Assay Buffer (prepare right before adding to Substrate Blank(s) in step 4).
  4. Mix 60 μL of 20 µg/mL rhDPEP1 and 60 μL 4 mM Substrate for a final concentration of 10 µg/mL and 2 mM respectively. Include a Substrate Blank containing 60 μL of 20 µg/mL rhDPEP1, 60 μL 1% TCA added 1 minute prior to substrate addition, and 60 μL Substrate.
  5. Incubate reactions for 1 hour at 37 °C.
  6. Dilute 10% TCA to 1% in Assay Buffer (prepare right before adding to reactions in step 7).
  7. Stop reactions by adding 60 μL of 1% TCA (do not add to Substrate Blank(s)) and incubate for 10 minutes at room temperature.
  8. Dilute OPA to 5 mg/mL in 2 M NaOH.
  9. Add 60 μL of 5 mg/mL OPA to each reaction.
  10. Incubate for 1 hour at room temperature.
  11. Load 200 µL of each incubated sample into the plate.
  12. Read at excitation and emission wavelengths of 360 nm and 460 nm (top read), respectively, in endpoint mode.
  13. Calculate Specific Activity:

     Specific Activity (pmol/min/µg) =

Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

   

  *Adjusted for Substrate Blank
     **Derived using calibration standard L-Histidine.
Per Well:
  • rhDPEP1: 1.0 µg
  • Substrate: 1 mM















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