CD26/Dipeptidyl Peptidase 4 蛋白, Human (HEK293)

The CD26/dipeptidyl peptidase 4 protein is a cell surface glycoprotein receptor that induces T cell proliferation and NF-kappa-B activation. CD26 also works with FAP to promote pericellular proteolysis, endothelial cell migration, and invasion of the extracellular matrix. CD26/Dipeptidyl Peptidase 4 Protein, Human (HEK293) is the recombinant human-derived CD26/Dipeptidyl Peptidase 4 protein, expressed by HEK293 , with tag free.

CD26/二肽基肽酶 4 蛋白是一种细胞表面糖蛋白受体，在 T 细胞受体 (TCR) 介导的 T 细胞激活所必需的共刺激信号中发挥着关键作用。作为 T 细胞共激活的正调节因子，CD26 与 ADA、CAV1、IGF2R 和 PTPRC 相互作用，以 T 细胞受体/CD3 依赖性方式导致 T 细胞增殖和 NF-κ-B 激活。它与 CAV1 和 CARD11 的结合诱导 T 细胞增殖，而它与 ADA 的相互作用则调节淋巴细胞-上皮细胞粘附。CD26 与 FAP 相关，参与细胞外基质的细胞周蛋白水解，有助于内皮细胞的迁移和侵袭。此外，它可能在促进淋巴管内皮细胞粘附、迁移和管形成方面发挥作用。CD26 作为具有二肽基肽酶活性的丝氨酸外肽酶，通过切割循环中的肽来调节各种生理过程，包括趋化因子、促有丝分裂生长因子、神经肽和肽激素。值得注意的是，CD26 在微生物感染过程中充当人类冠状病毒 MERS-CoV-2 的受体。

Measured by its ability to cleave the fluorogenic peptide substrate Gly-Pro-7-amido-4-methylcoumarin (GP-AMC) and the specific activity is > 2,500 pmol/min/μg.

Materials
Assay buffer: 25 mM Tris, pH 8.0
CD26/Dipeptidyl Peptidase 4 Protein, Human (HEK293) (HY-P72975)
Substrate: H-Gly-Pro-AMC (HY-137834)
Standard: 7-Amino-4-methylcoumarin (AMC, HY-D0027)

Procedure
1. Standard Curve: Dilute standard in assay buffer to concentrations of 0, 0.09765625, 0.1953125, 0.390625, 0.78125, 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 μM. 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 Human CD26 to 0.2 μg/mL in assay buffer.
3. Dilute the substrate to 200 μM in assay buffer.
4. Add 50 μL of 0.2 μg/mL Human CD26 to the plate, then add 50 μL of 200 μM substrate to initiate the reaction. For the substrate blank, load 50 μL of assay buffer and 50 μL of substrate.
5. Read in kinetic mode for 5 minutes at excitation wavelength 380 nm and emission wavelength 460 nm.
6. Calculate the specific activity:

Human

HEK293

Tag Free

P27487 (N29-P766)

1803  [NCBI]

Extracellular Domain

NKGTDDATADSRKTYTLTDYLKNTYRLKLYSLRWISDHEYLYKQENNILVFNAEYGNSSVFLENSTFDEFGHSINDYSISPDGQFILLEYNYVKQWRHSYTASYDIYDLNKRQLITEERIPNNTQWVTWSPVGHKLAYVWNNDIYVKIEPNLPSYRITWTGKEDIIYNGITDWVYEEEVFSAYSALWWSPNGTFLAYAQFNDTEVPLIEYSFYSDESLQYPKTVRVPYPKAGAVNPTVKFFVVNTDSLSSVTNATSIQITAPASMLIGDHYLCDVTWATQERISLQWLRRIQNYSVMDICDYDESSGRWNCLVARQHIEMSTTGWVGRFRPSEPHFTLDGNSFYKIISNEEGYRHICYFQIDKKDCTFITKGTWEVIGIEALTSDYLYYISNEYKGMPGGRNLYKIQLSDYTKVTCLSCELNPERCQYYSVSFSKEAKYYQLRCSGPGLPLYTLHSSVNDKGLRVLEDNSALDKMLQNVQMPSKKLDFIILNETKFWYQMILPPHFDKSKKYPLLLDVYAGPCSQKADTVFRLNWATYLASTENIIVASFDGRGSGYQGDKIMHAINRRLGTFEVEDQIEAARQFSKMGFVDNKRIAIWGWSYGGYVTSMVLGSGSGVFKCGIAVAPVSRWEYYDSVYTERYMGLPTPEDNLDHYRNSTVMSRAENFKQVEYLLIHGTADDNVHFQQSAQISKALVDVGVDFQAMWYTDEDHGIASSTAHQHIYTHMSHFIKQCFSLP

Approximately 90-130 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

无菌粉末

Lyophilized from a 0.2 μm filtered solution of 25 mM MES, 150 mM NaCl, pH 6.5, 5% Trehalose, 5% Mannitol, 0.01% Tween-80 or 50 mM HEPES, 150 mM NaCl, pH 7.5, 8% trehalose.

<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.