FAP Protein, Mouse (HEK293, His)

FAP protein is a cell surface glycoprotein serine protease that regulates tissue remodeling, fibrosis, inflammation, and tumor growth. It cleaves post-proline residues, degrades SERPINF2 and SPRY2, and exhibits dipeptidyl peptidase activity. FAP Protein, Mouse (HEK293, His) is the recombinant mouse-derived FAP protein, expressed by HEK293 , with N-His labeled tag.

FAP Protein is a cell surface glycoprotein serine protease that is involved in various cellular processes such as tissue remodeling, fibrosis, wound healing, inflammation, and tumor growth. It exhibits endopeptidase activity, particularly cleaving post-proline residues, and can degrade substances like alpha-2-antiplasmin SERPINF2 and SPRY2. FAP Protein also has dipeptidyl peptidase activity, with a preference for specific dipeptide substrates. It interacts with other proteins like DPP4, PLAUR, and integrins to promote pericellular proteolysis of the extracellular matrix, leading to cell adhesion, migration, and invasion. Additionally, FAP Protein plays a role in tissue remodeling, wound healing, and cell invasiveness in malignant melanoma cancers. It also enhances tumor growth by promoting angiogenesis, collagen fiber degradation, and apoptosis while reducing the immune system's antitumor response. In melanocytic cells, FAP Protein acts as a tumor suppressor by regulating cell proliferation and survival independently of its serine protease activity.

1.Measured by its ability to convert the substrate benzyloxycarbonyl-Gly-Pro-7-amido-4-methylcoumarin (Z-GP-AMC) to Z-Gly-Pro and 7-amino-4-methylcoumarin (AMC). The specific activity is >2000 pmol/min/µg.
2.Immobilized Mouse FAP, His Tag at 1 μg/mL (100 μl/well) on the plate. Dose response curve for Anti-FAP Antibody, hFc Tag with the EC₅₀ of 4.6 ng/mL determined by ELISA.

Materials
Assay buffer: 50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
Test protein: FAP Protein, Mouse (HEK293, His) (HY-P77934)
Substrate: Dissolve Z-Gly-Pro-AMC (HY-D1670) in DMSO to a concentration of 10 mM.
Standard: 7-Amino, 4-Methyl Coumarin (HY-D0027)

Procedure
1. Standard Curve: Dilute standard in assay buffer to concentrations of 0, 0.78, 1.5625, 3.125, 6.25, 12.5, 25, and 50 μmol/L. Add 100 μL of each dilution to a black microplate well. Measure fluorescence in kinetic mode for 5 minutes at excitation and emission wavelengths of 380 nm and 460 nm, respectively. Plot the measured RFU values on the y-axis and the standard concentrations on the x-axis to generate a standard curve and obtain the curve equation.
2. Dilute Mouse FAP to 0.2 μg/mL in assay buffer.
3. Dilute the substrate to 100 μM in assay buffer.
Experimental group: Add 50 μL of 2 μg/mL FAP to the plate and initiate the reaction by adding 50 μL of 100 μM substrate.
Substrate blank group: 50 μL assay buffer + 50 μL, 100 μM substrate.
4. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 380 nm and 460 nm, respectively.
5. Calculate Specific Activity:

Mouse

HEK293

N-His

P97321-1 (L26-D761)

14089  [NCBI]

Extracellular Domain

LRPSRVYKPEGNTKRALTLKDILNGTFSYKTYFPNWISEQEYLHQSEDDNIVFYNIETRESYIILSNSTMKSVNATDYGLSPDRQFVYLESDYSKLWRYSYTATYYIYDLQNGEFVRGYELPRPIQYLCWSPVGSKLAYVYQNNIYLKQRPGDPPFQITYTGRENRIFNGIPDWVYEEEMLATKYALWWSPDGKFLAYVEFNDSDIPIIAYSYYGDGQYPRTINIPYPKAGAKNPVVRVFIVDTTYPHHVGPMEVPVPEMIASSDYYFSWLTWVSSERVCLQWLKRVQNVSVLSICDFREDWHAWECPKNQEHVEESRTGWAGGFFVSTPAFSQDATSYYKIFSDKDGYKHIHYIKDTVENAIQITSGKWEAIYIFRVTQDSLFYSSNEFEGYPGRRNIYRISIGNSPPSKKCVTCHLRKERCQYYTASFSYKAKYYALVCYGPGLPISTLHDGRTDQEIQVLEENKELENSLRNIQLPKVEIKKLKDGGLTFWYKMILPPQFDRSKKYPLLIQVYGGPCSQSVKSVFAVNWITYLASKEGIVIALVDGRGTAFQGDKFLHAVYRKLGVYEVEDQLTAVRKFIEMGFIDEERIAIWGWSYGGYVSSLALASGTGLFKCGIAVAPVSSWEYYASIYSERFMGLPTKDDNLEHYKNSTVMARAEYFRNVDYLLIHGTADDNVHFQNSAQIAKALVNAQVDFQAMWYSDQNHGISSGRSQNHLYTHMTHFLKQCFSLSD

Approximately 90-110 kDa due to the glycosylation

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder

1.Lyophilized from a 0.22 μm filtered solution of 50 mM Tris-HCl, 300 mM NaCl, pH 8.0.
2.Lyophilized from a 0.22 μm filtered solution of PBS, 8% trehalose, pH 7.4.
3.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
4.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.
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.

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.

• [1]. /biology-dictionary/baf3-cell.html  [Content Brief]