Recombinant Human Lactate Dehydrogenase A/LDHA Protein, CF

R&D Systems | Catalog # 9158-HA

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Lactate Dehydrogenase A/LDHA Protein (9158-HA)
  • 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

P00338

Applications

Enzyme Activity
View all Lactate Dehydrogenase A/LDHA Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Lactate Dehydrogenase A/LDHA protein
Ala2-Phe332, with N-terminal Met and 6-His tag

Purity

>95%, 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

Met

Predicted Molecular Mass

38 kDa

SDS-PAGE

36 kDa, reducing conditions

Activity

Measured by its ability to reduce pyruvate to lactate.
The specific activity is >95,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

9158-HA
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: Lactate Dehydrogenase A/LDHA

A hallmark of most cancer cells is an altered metabolism involving a shift to aerobic glycolysis with lactate production coupled with a higher uptake of glucose as the main source of energy. Lactate dehydrogenase (LDH) is key to this shift by catalyzing the formation of lactate by reducing pyruvate with NADH, which also generates NAD(+) that is essential for the continuity of glycolysis (1). LDHA is a key enzyme that controls the production of lactate in the glycolysis pathway. It is therefore an important control point in the system of cellular energy release. It's up regulation is common in many malignant tumors. Inhibiting LDH activity has an anti-proliferative effect on cancer cells (2). It may reverse the resistance of tumor cells to conventional chemo- and radiotherapy. Recent research has renewed interest in LDH as an anticancer drug target (3).The protein is found predominantly in muscle tissue. Mutations in LDHA have been linked to exertional myoglobinuria (4).

References

  1. Faloppi L. et al. (2016) Sci Rep. doi: 10.1038/srep24136.
  2. Ghosh, M. et al. (2016) Chem. Commun. 52:2401.
  3. Augoff, K. et al. (2015) Cancer Lett. 358:1.
  4. Maekawa M. et al. (1990). Biochem. Biophys. Res. Commun. 168:677.

Alternate Names

LDH1, LDHA, LDHM

Entrez Gene IDs

3939 (Human); 16828 (Mouse); 24533 (Rat)

Gene Symbol

LDHA

UniProt

P00338 (Human)

Additional Lactate Dehydrogenase A/LDHA Products

Product Documents for Recombinant Human Lactate Dehydrogenase A/LDHA 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 Lactate Dehydrogenase A/LDHA Protein, CF

For research use only

Customer Reviews for Recombinant Human Lactate Dehydrogenase A/LDHA Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Lactate Dehydrogenase A/LDHA Protein, CF and earn rewards!

Have you used Recombinant Human Lactate Dehydrogenase A/LDHA 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 Lactate Dehydrogenase A/LDHA Protein, CF (9158-HA):

Materials
  •  Assay Buffer:  25 mM Tris, 100 mM NaCl, pH 7.5
  • Recombinant Human Lactate Dehydrogenase A/LDHA (rhLDHA) (Catalog # 9158-HA)
  • 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
  • 100 mM Sodium Pyruvate (Gibco, Catalog # 11360)
  • 96-well Clear Plate  (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhLDHA to 0.4 ng/µL in Assay Buffer.
  2. Prepare a substrate mixture containing 1.6 mM beta -NADH and 4 mM sodium pyruvate in Assay Buffer.
  3. In a plate load 50 µL of 0.4 ng/µL rhLDHA, and start the reaction by adding 50 µL of substrate mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of substrate mixture.
  4. Read plate at 340 nm (absorbance) in kinetic mode for 5 minutes.
  5. 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 Substrate Blank.
     **Using the extinction coefficient 6220 M-1cm-1.
     ***Using the path correction 0.320 cm.
     Note: the output of many spectrophotometers is in mOD.

Per Well:

  • rhLDHA: 0.020 µg
  • beta -NADH: 0.8 mM
  • Sodium pyruvate: 2 mM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes