Recombinant Human NM23-H1 His-tag Protein, CF

R&D Systems | Catalog # 10495-NM

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human NM23-H1 His-tag Protein (10495-NM)
  • 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

E. coli

Accession Number

P15531.1

Applications

Enzyme Activity
View all NM23-H1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human NM23-H1 protein
Ala2-Glu152
with a 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

Ala2

Predicted Molecular Mass

18 kDa

SDS-PAGE

18-20 kDa, under reducing conditions.

Activity

Measured by its ability to convert thymidine diphosphate and adenosine triphosphate to thymidine triphosphate and adenosine diphosphate in a coupled assay.
The specific activity is >50,000 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human NM23-H1 His-tag Protein, CF

Recombinant Human NM23-H1 His-tag Protein Enzyme Activity

Recombinant Human NM23-H1 His-tag Protein Enzyme Activity

Recombinant Human NM23-H1 His-tag (Catalog # 10495-NM) is measured by its ability to convert thymidine diphosphate and adenosine triphosphate to thymidine triphosphate and adenosine diphosphate in a coupled assay.

Recombinant Human NM23-H1 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human NM23-H1 His-tag Protein (Catalog # 10495-NM) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 18-20 kDa.

Formulation, Preparation, and Storage

10495-NM
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and TCEP.
Shipping The product is shipped with dry ice or equivalent. 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: NM23-H1

Non-metastatic Protein 23 Homolog 1 (NM23-H1), also known as Nucleoside diphosphate kinase A (NDPK-A), is a group 1 member of the NDPK family encoded by the NME1 (non metastatic cell) gene (1). NDPKs are well-known as housekeeping genes that maintain the balance of nucleoside triphosphate levels by transferring a phosphate group from a nucleoside triphosphate to nucleoside diphosphate. NM23-H1 is a cytosolic protein that forms active hexamers composed of a trimer of dimers (2,3). The active hexamer contains a "head" region with substrate binding and catalytic activity as well as a Kpn loop and C-terminal extensions that stabilize the structure (2,3). NM23-H1 (NDPK-A) shares significant homology with NM23-H2 (NDPK-B), with 88% identity (3). In spite of significant homology and catalytic function between these isoforms, the differing amino acids on the lateral surface of the hexamers lead to specific interactions with other proteins and ultimately result in diverging cellular distribution and function in cells (1). NM23-H1 was the first reported metastasis suppressor gene (4) and plays a role in cell adhesion, migration and motility, and signaling and proteolysis (5-7). Suppression activity has been demonstrated in many cancers although the mechanism involved is uncertain (1,7). NM23-H2 has been reported to have several functions: enzymatic function as a protein histidine kinase with ATP-citrate lyase and annexin A1 as direct targets (5), exonuclease activity (8), NDPK activity (1), and also forms a complex with the cystic fibrosis transmembrane conductance regulator (CFTR) (9) in regulation of chloride channels, other signaling pathways, and gene regulation (1,10,11).

References

  1. Boissan, M. et al. (2018) Lab. Invest. 98:164.
  2. Janin, J. et al. (2000) J. Bioenerg. Biomembr. 32:215.
  3. Webb, P.A. et al. (1995) J. Mol. Biol. 251:574.
  4. Steeg, P.S. et al. (1988) J. Natl. Cancer Inst. 80:200.
  5. Attwood, P.V. et al. (2018) Lab. Invest. 98:283.
  6. Khan, I. et al. (2019) Cancer Res. 79:4689.
  7. Matyasi, B. et al. (2020) Pathol. Oncol. Res. 26:49.
  8. Fan, Z. et al. (2003) Cell 112:659.
  9. Borthwick, L.A. et al. (2016) PloS One. 11:e0149097.
  10. Ma, W. et al. (2008) Biochem. Biophys. Res. Commun. 371:425.
  11. Zheng, S. et al. (2019) FEBS Lett. 593:80.

Long Name

Non-metastatic Protein 23 Homolog 1

Alternate Names

GAAD, NDKA, NDPKA, NM23A, NM23H1, NME1

Entrez Gene IDs

4830 (Human)

Gene Symbol

NME1

UniProt

P15531.1

Additional NM23-H1 Products

Product Documents for Recombinant Human NM23-H1 His-tag 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 NM23-H1 His-tag Protein, CF

For research use only

Related Research Areas

Cancer Biomarkers

Customer Reviews for Recombinant Human NM23-H1 His-tag Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human NM23-H1 His-tag Protein, CF and earn rewards!

Have you used Recombinant Human NM23-H1 His-tag Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human NM23-H1 His-tag Protein, CF (10495-NM):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM MgCl2, 100 mM KCl, pH 7.5
  • Recombinant Human NM23-H1 His-tag (rhNM23-H1) (Catalog # 10495-NM)
  • Substrate: Thymidine 5'-Diphosphate (TDP) (Sigma, Catalog # T9375), 40 mM stock in 10 mM Sodium Borate, pH 9.0
  • Recombinant Human PKM2 (rhPKM2) (Catalog # 7244-PK)
  • Recombinant Human Lactate Dehydrogenase A/LDHA (rhLDHA) (Catalog # 9158-HA)
  • Adenosine 5'-triphosphate (ATP) (Sigma, Catalog # A7699), 400 mM stock in deionized water
  • beta -Nicotinamide adenine dinucleotide, reduced disodium salt hydrate ( beta -NADH) (Sigma, Catalog # N8129), 20 mM stock in 0.1 M Sodium Borate, pH 9.0
  • Phospho(enol)pyruvic acid (PEP) (Sigma, Catalog # P0564), 50 mM stock in deionized water
  • 96-well clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare Reaction Mixture containing 20 µg/mL rhPKM2, 5 µg/mL rhLDHA, 4 mM ATP, 400 µM beta -NADH, 2 mM PEP and 2 mM TDP in Assay Buffer.
  2. Incubate Reaction Mixture at room temperature for 5 minutes.
  3. Dilute rhNM23-H1 to 0.01 ng/µL in Assay Buffer.
  4. In a plate, load 50 µL of 0.01 ng/µL rhNM23-H1 and start the reaction by adding 50 µL of Reaction Mixture. Include a Control containing 50 µL of Assay Buffer and 50 µL of Reaction Mixture.
  5. Read at an absorbance of 340 nm in kinetic mode for 5 minutes.
  6. Calculate specific activity:

         Specific Activity (pmol/min/µg) =

    		Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol x (-1)
    ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

*Adjusted for Control
**Using the extinction coefficient 6220 M-1cm-1
***Using the path correction 0.32 cm
Note: the output of many spectrophotometers is in mOD

Per Well:

  • rhNM23-H1: 0.0005 µg
  • TDP: 1 mM
  • ATP: 2 mM
  • beta -NADH: 200 µM
  • PEP: 1 mM
  • rhPKM2: 1 µg
  • rhLDHA: 0.25 µg

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes