ADAM9 Protein, Human (HEK293, His, solution)

ADAM9, a metalloprotease, cleaves and releases TEK, KDR, EPHB4, CD40, VCAM1, and CDH5, playing a pivotal role in tumorigenesis and angiogenesis. It regulates cell motility through integrin interactions, mediates cell-cell and cell-matrix interactions, and may act as an alpha-secretase for APP, suggesting involvement in neurobiology and neurodegenerative disorders. ADAM9 Protein, Human (HEK293, His, solution) is the recombinant human-derived ADAM9 protein, expressed by HEK293 , with C-His labeled tag.

ADAM9, a metalloprotease, plays a pivotal role in tumorigenesis and angiogenesis by cleaving and releasing various molecules including TEK, KDR, EPHB4, CD40, VCAM1, and CDH5. Its multifaceted functions extend to mediating cell-cell and cell-matrix interactions, thereby regulating cell motility through interactions with integrins. Additionally, ADAM9 may function as an alpha-secretase for the amyloid precursor protein (APP), highlighting its involvement in processes relevant to neurobiology and neurodegenerative disorders.

Measured by its ability to cleave a fluorogenic peptidesubstrate Mca-PLAQAV-Dpa-RSSSR-NH. The specificactivity is 6.85 pmol/min/ug.

Materials
Assay buffer: 25 mM Tris, 2.5 μM ZnCl₂, 0.005% (w/v) Brij-35, pH 9.0
ADAM9 Protein, Human (HEK293, His, Solution) (HY-P77867)
Substrate: Mca-PLAQAV-Dpa-RSSSR-NH₂ TFA (HY-P3722A)
Standard: MCA-Pro-Leu-OH

Procedure
1. Dilute MCA-Pro-Leu-OH to 50, 25, 12.5, 6.25, 3.125, 1.5625, 0.78125, and 0 μmol/L respectively with assay buffer. Add 100 μL of each dilution to the wells to initiate the reaction. Set the excitation and emission wavelengths to 320 nm and 405 nm respectively, and read in kinetic mode for 5 minutes. Plot a standard curve with the standard concentration as the abscissa and the measured RFU as the ordinate, and obtain the standard curve equation.
2. Dilute Human ADAM9 to 40 μg/mL in assay buffer.
3. Dilute the substrate to 20 μM in assay buffer.
4. Mix equal volumes of 40 μg/mL Human ADAM9 and 20 μM substrate, including controls containing assay buffer and 20 μM substrate without any Human ADAM9.
5. Incubate the reaction mixture at 37°C for 30 minutes.
6. Transfer 100 μL from each well to a black plate.
7. Read in endpoint mode using excitation and emission wavelengths of 320 nm and 405 nm respectively.
8. Calculate the specific activity:

Human

HEK293

C-8*His

Q13443-1 (A29-D697)

8754  [NCBI]

Extracellular Domain

40-45 kDa and 60-70 kDa

Greater than 95% as determined by Tris-Bis PAGE.

Solution.

Supplied as a 0.22 μm filtered solution of 50 mM Tris-HCl, 500 mM NaCl, 0.005% Brij-35,20% Glycerol, pH 8.5.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

N/A.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.