Recombinant Mouse TACE/ADAM17 Protein, CF

Catalog #: 2978-AD
5 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
2978-AD-010

R&D Systems Recombinant Proteins and Enzymes

1 Image

Product Details

Citations (5)

Supplemental Products

Reviews

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse TACE/ADAM17 Protein, CF Summary

Product Specifications

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

Activity

Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLAQAV-Dpa-RSSSR-NH₂ (Catalog # ES003). The specific activity is >500 pmol/min/µg, as measured under the described conditions. Note: It is extremely important that the assay solution does not contain salt as it inhibits TACE activity.

Source

Mouse myeloma cell line, NS0-derived mouse TACE/ADAM17 protein
Pro18-Asp563 and Arg215-Asp563, both with a C-terminal 10-His tag, Pro18-Arg214 and Asp59-Arg214

Accession #

AAC62934

N-terminal Sequence
Analysis

Pro18, Asp59 & Arg215

Structure / Form

Recombinant Mouse TACE/ADAM17 is prone to proteolytic cleavage at C-terminus. The poly-His tag may not be present in the preparation.

Predicted Molecular Mass

63 kDa, 40 kDa, 23 kDa, 18 kDa

SDS-PAGE

76 kDa, 63 kDa, 21 kDa and 18 kDa, reducing conditions

Product Datasheets

2978-AD

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

2978-AD

Formulation	Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Reconstitution	Reconstitute at 100 μg/mL in sterile, deionized water.
Shipping	The product is shipped at ambient temperature. 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 reconstitution.

Assay Procedure

Materials

Assay Buffer: 50 mM Tris, pH 9.0
Recombinant Mouse TACE/ADAM17 (rmTACE) (Catalog # 2978-AD)
Substrate: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH₂ (Catalog # ES003), 2 mM stock in DMSO
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

Dilute rmTACE to 0.2 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
In a plate, load 50 µL of 0.2 ng/µL rmTACE to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank of 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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:

rmTACE: 0.01 µg
Substrate: 10 µM

Reconstitution Calculator

Background: TACE/ADAM17

Tumor necrosis factor alpha (TNF-alpha ) converting enzyme (TACE) is a member of the ADAM family that contains a disintegrin and metalloprotease-like domain. The domain structure of mouse TACE consists of pro (aa 18-214), catalytic (aa 215-474), disintegrin-like (aa 475-563), cysteine-rich (aa 564-602), Crambin-like (aa 603-671), transmembrane (aa 672-692), and cytoplasmic (aa 693-827) (1, 2). Recombinant mouse TACE ends after the disintegrin-like domain and consists of several forms as described. In addition to its ability to release the 17 kDa extracellular form of TNF-alpha from the 26 kDa membrane-anchored TNF-alpha, TACE also plays an essential role in shedding ectodomains from a variety of proteins such as L-selectin, transforming growth factor-alpha, amyloid protein precursor, and Notch1 receptor. TACE mRNA is present in virtually every tissue and TACE protein resides both on the cell surface and in the cell (3).

References

Cerretti, D.P. et al. (1999) Cytokine 11:541.
Mizui, Y. et al. (1999) Gene 233:67.
Black, R.A. (2004) Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. pp. 726, Academic Press, San Diego.

Long Name

TNF-alpha Converting Enzyme

Entrez Gene IDs

6868 (Human); 11491 (Mouse)

Alternate Names

ADAM 17; ADAM metallopeptidase domain 17; ADAM metallopeptidase domain 18; ADAM17; ADAM18; CD156b antigen; CD156b; CSVP; disintegrin and metalloproteinase domain-containing protein 17; EC 3.4.24.86; MGC71942; Snake venom-like protease; TACE; TACEcSVP; TNF-alpha convertase; TNF-alpha converting enzyme; TNF-alpha-converting enzyme; tumor necrosis factor, alpha, converting enzyme

Related Research Areas

Citations for Recombinant Mouse TACE/ADAM17 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
Filter your results:

Filter by: