ADAM9 Protein, Mouse (HEK293, His)

ADAM9 protein is a metalloprotease that plays an important role in tumorigenesis and angiogenesis. It cleaves TEK, KDR, EPHB4, CD40, VCAM1 and CDH5, regulating cancer-related signaling pathways and blood vessel formation. ADAM9 Protein, Mouse (HEK293, His) is the recombinant mouse-derived ADAM9 protein, expressed by HEK293 , with C-His labeled tag.

ADAM9 Protein, a metalloprotease, is implicated in crucial cellular processes associated with tumorigenesis and angiogenesis. This multifaceted enzyme is known to cleave and release various molecules, including TEK, KDR, EPHB4, CD40, VCAM1, and CDH5, highlighting its involvement in modulating key signaling pathways relevant to cancer progression and blood vessel formation. Beyond its role in proteolytic events, ADAM9 is suggested to play a part in mediating cell-cell and cell-matrix interactions, thereby contributing to the regulation of cellular motility, potentially through interactions with integrins. This diverse range of molecular interactions underscores the significance of ADAM9 in influencing cellular behavior and its potential impact on pathological processes.

Measured by its ability to cleave a fluorogenic peptidesubstrate Mca-PLAQAV-Dpa-RSSSR-NH₂. The specificactivity is 1.72 pmol/min/ug.

Materials
Assay Buffer: Assay Buffer: 25 mM Tris, 2.5 µM ZnCl2, 0.005% (w/v) Brij-35, pH 9.0
Recombinant Human ADAM-9 (rhADAM9)
Fluorogenic Peptide Substrate III: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH₂
F16 Black Maxisorp Plate
Fluorescent Plate Reader

1.Dilute rhADAM9 to 40 µg/mL in Assay Buffer.
2.Dilute substrate to 20 µM in Assay Buffer.
3.Combine equal volumes of 40 µg/mL rhADAM9 and 20 µM substrate. Include a control containing Assay Buffer and 20 µM substrate without any rhADAM9.
4.Incubate reactions at 37 °C for 30 minutes.
5.Load 100 μL of reactions and control per well in a black plate.
6.Read with excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in endpoint mode.
7.Calculate specific activity:

Mouse

HEK293

C-His

Q61072 (G30-D697)

11502  [NCBI]

Extracellular Domain

GRPDLEQTVHLSSYEIITPWRLTRERREALGPSSQQISYVIQAQGKQHIIHLERNTDLLPNDFVVYTYDKEGSLLSDHPNVQSHCHYRGYVEGVQNSAVAVSACFGLRGLLHLENASFGIEPLHNSSHFEHIFYPMDGIHQEPLRCGVSNRDTEKEGTQGDEEEHPSVTQLLRRRRAVLPQTRYVELFIVVDKERYDMMGRNQTAVREEMIRLANYLDSMYIMLNIRIVLVGLEIWTDRNPINIIGGAGDVLGNFVQWREKFLITRWRHDSAQLVLKKGFGGTAGMAFVGTVCSRSHAGGINVFGQITVETFASIVAHELGHNLGMNHDDGRECFCGAKSCIMNSGASGSRNFSSCSAEDFEKLTLNKGGSCLLNIPKPDEAYSAPSCGNKLVDPGEECDCGTAKECEVDPCCEGSTCKLKSFAECAYGDCCKDCQFLPGGSMCRGKTSECDVPEYCNGSSQFCPPDVFIQNGYPCQNSKAYCYNGMCQYYDAQCQVIFGSKAKAAPRDCFIEVNSKGDRFGNCGFSGSEYKKCATGNALCGKLQCENVQDMPVFGIVPAIIQTPSRGTKCWGVDFQLGSDVPDPGMVNEGTKCDAGKICRNFQCVNASVLNYDCDIQGKCHGHGVCNSNKNCHCEDGWAPPHCDTKGYGGSVDSGPTYNAKSTALRD

Predicted band size: 77.61 kDa; Observed band size: 65-115 kDa

Greater than 90% as determined by reducing SDS-PAGE

Lyophilized powder

Lyophilized from a 0.22 μm filtered solution of PBS (pH 7.4). Normally 8% trehalose is added as protectant before lyophilization.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.