Recombinant Human Napsin A Protein, CF

R&D Systems | Catalog # 8489-NA

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Napsin A Protein (8489-NA)
  • 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

O96009

Applications

Enzyme Activity
View all Napsin A Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Napsin A protein
Thr25-Gly420, with C-terminal 6-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

Thr25

Predicted Molecular Mass

44 kDa

SDS-PAGE

40-70 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).
The specific activity is >12 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

8489-NA
Formulation Supplied as a 0.2 μm filtered solution in MES 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: Napsin A

Human NAPSA (also called Napsin A) is a 38 kDa glycosylated aspartic protease (1-3). NAPSA/Napsin A is synthesized as a 420 amino acid (aa) proenzyme containing a signal peptide (up to 25 aa), an activation peptide (38 aa), and a 357 aa mature enzyme. Mature human NAPSA/Napsin A shares 74% and 73% aa sequence identity with mature mouse and rat NAPSA/Napsin A, respectively. The NAPSA/Napsin A proenzyme is expressed in lung epithelial cells and within the kidney (3). Kidney dysfunction is associated with a decrease in NAPSA/Napsin A excretion in urine (1). NAPSA/Napsin A expression is also used as a marker for clear cell ovarian carcinomas, pulmonary adenocarcinomas, and renal cell carcinomas (4-8). NAPSA/Napsin A is involved in processing surfactant protein B in type II pneumocytes (9).

References

  1. Schauer-Vukasinovic, V. et al. (2001) Biochim. Biophys. Acta 1524:51.
  2. Tatnell, P.J. et al. (1998) FEBS Lett. 441:43.
  3. Schauer-Vukasinovic, V. et al. (1999) FEBS Lett. 462:135.
  4. Yamashita, Y. et al. (2014) Mod. Pathol. PMID 24721826
  5. Skirnisdottir, I. et al. (2013) BMC Cancer. 13:524.
  6. Ordonez, N.G. (2012) Adv. Anat. Pathol. 19:66.
  7. Jagirdar, J. (2008) Arch. Pathol. Lab. Med. 132:384.
  8. Ao, M.H. et al. (2014) Hum. Pathol. 45:926.
  9. Brasch, F. et al. (2003) J. Biol. Chem. 278:49006.

Alternate Names

ASP4, KAP, NAPA, NAPSA, Napsin-1, SNAPA, TA01/TA02

Entrez Gene IDs

9476 (Human); 16541 (Mouse); 60575 (Rat)

Gene Symbol

NAPSA

UniProt

O96009

Additional Napsin A Products

Product Documents for Recombinant Human Napsin A 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.

Product Specific Notices for Recombinant Human Napsin A Protein, CF

For research use only

Related Research Areas

Cancer Biomarkers
Lysosomal Enzymes

Citations for Recombinant Human Napsin A Protein, CF

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Protocols

View specific protocols for Recombinant Human Napsin A Protein, CF (8489-NA):

Materials
  • Assay Buffer: 0.1 M NaOAc, 0.2 M NaCl, pH 3.5
  • Recombinant Human Napsin A (rhNAPSA) (Catalog # 8489-NA)
  • Substrate: MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Catalog # ES001), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhNAPSA to 40 µg/mL in Assay Buffer.
  2. Incubate 50 µL aliquot of 40 µg/mL rhNAPSA at room temperature for 20 minutes.
  3. Dilute incubated rhNAPSA to 8 µg/mL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load 50 µL incubated rhNAPSA into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 µL of 20 µM Substrate.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     
      *Adjusted for Substrate Blank

      **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • rhNAPSA: 0.4 µg
  • Substrate: 10 µM

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