The DPPIV Drug Discovery Kit is a complete assay system designed to screen DPPIV inhibitors, providing enough material to perform at least 96 assays. DPPIV (DPP4, CD26) is a member of the class of proteases known as prolyl peptidases, which cleave proteins after proline residues ａnd is thought to play roles in diabetes, cancer, ａnd autoimmune diseases, making it a target for drug discovery.

 

The kit contains both a chromogenic substrate (H-Gly-Pro-pNA; Km=114 ?M) ａnd a fluorogenic substrate (H-Gly-Pro-AMC; Km=50 ?M).

 

Cleavage of the p-nitroaniline (pNA) from the colorimetric substrate increases absorbance at 405 nm. The fluorimetric assay is based on the cleavage of 7-amino-4-methylcoumarin (AMC) moiety from the C-terminus of the peptide substrate, which increases its fluorescence intensity at 460 nm. The kit is useful to screen inhibitors of DPPIV, a potential therapeutic target. A DPPIV inhibitor, P32/98 (KI=130 nM12), is included for use as a control. Other DPP enzymes are available for specificity profiling.

Product Specification

Alternative Name:CD26 drug discovery kit, Dipeptidyl Peptidase IV drug discovery kit, DPP4 drug discovery kit

 

Long Term Storage:-80°C

 

Kit/Set Contains:DPPIV Enzyme (Human, recombinant) (Prod. No. BML-SE434-9090) (35 mU (40.4?l); 0.867 mU/?l. On U=1?mole/min@37°C, 100?M H-Gly-Pro-pNA. Purity ≥95% (SDS-Page)) Storage: -70°C; avoid freeze thaw cycles

pNA substrate (Prod. No. BML-PI188-9090) (150?l; 10mM in DMSO) Storage: -70°C

Calibration standard (p-nitroaniline) (Prod. No. BML-KI106) (1ml; 50?M in assay buffer) Storage: -70°C

AMC Substrate (Prod. No. BML-P189-9090) (150?l; 0.5mM in DMSO) Storage: -70°C

 

Calibration standard (7-Amino-4-methylcoumarin) (Prod. No. BML-KI107) (1ml; 40?M in assay buffer) Storage: -70°C)

Inhibitor (Prod. No. BML-PI142-9090) (20?l; 1mM in DMSO) Storage: -70°C

Assay Buffer (50mM TRIS, pH 7.5) (Prod. No. BML-KI342) (20ml; liquid in screw-cap plastic bottle) Storage: -20 or -70°C

?-volume clear microplate (Prod. No. BML-K101)Storage: Room temperature

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Product Literature References

16, 17-Dihydro-17b-hydroxy isomitraphylline alkaloid as an inhibitor of DPP-IV, ａnd its effect on incretin hormone ａnd β-cell proliferation in diabetic rat: A. Shukla, et al.; Eur. J. Pharm. 47, 512 (2012), Application(s): Measurement of inhibition of DPPIV by DHIM, Abstract;

Discovery ａnd preclinical profile of Saxagliptin (BMS-477118): a highly potent, long-acting, orally active dipeptidyl peptidase IV inhibitor for the treatment of type 2 diabetes: D. J. Augeri et al.; J. Med. Chem. 48, 5025 (2005), Abstract;

CD26/dipeptidyl peptidase IV ａnd its role in cancer: B. et al.; Histol. Histopathol. 19, 1345 (2004), Abstract;

CD26/dipeptidyl peptidase IV: a regulator of immune function ａnd a potential molecular target for therapy: U. Aytac et al.; Curr. Drug. Targets Immune Endocr. Metabol. Disord. 4, 11 (2004), Abstract;

Dipeptidyl peptidase IV inhibitors for the treatment of diabetes: A. E. Weber et al.; J. Med. Chem. 47, 4135 (2004),Abstract;

N-linked glycosylation of dipeptidyl peptidase IV (CD26): effects on enzyme activity, homodimer formation, ａnd adenosine deaminase binding: K. Aertgeerts et al.; Protein Sci. 13, 145 (2004), Abstract;

Dipeptidyl peptidase IV (CD26) activity in the hematopoietic system: differences between the membrane-anchored ａnd the released enzyme activity: D. A. Pereira et al.; Braz. J. Med. Biol. Res. 36, 567 (2003), Abstract;

Dipeptidyl peptidase IV inhibitor treatment stimulates beta-cell survival ａnd islet neogenesis in streptozotocin-induced diabetic rats: J. A. Pospisilik et al.; Diabetes 52, 741 (2003), Abstract;

Prolyl peptidases: a serine protease subfamily with high potential for drug discovery: J. S. Rosenblum et al.; Curr. Opin. Chem. Biol. 7, 496 (2003), Abstract;

Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV: R. Thoma et al.; Structure 11, 947 (2003), Abstract;

On the regulatory role of dipeptidyl peptidase IV (=CD=adenosine deaminase complexing protein) on adenosine deaminase activity: I. Ben-Shooshan; Biochim. Biophys. Acta. 1587, 21 (2002), Abstract;

Human serum dipeptidyl peptidase IV (DPPIV) ａnd its unique properties: H. Shibuya-Saruta; J. Clin. Lab. Anal. 10, 435 (1996), Abstract;

Activity of dipeptidyl peptidase IV ａnd post-proline cleaving enzyme in sera from osteoporotic patients: H. Gotoh et al.; Clin. Chem. 34, 2499 (1988), Abstract;

 

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二肽基肽酶(dipeptidyl peptidase ，DPP)

二肽基肽酶(dipeptidyl peptidase ，DPP)是蛋白酶家族中的脯基肽酶成员，其可剪切含脯氨酸的蛋白，其中DPPIV (DPP4, CD26),是一种丝氨酸二肽基肽酶，其剪切靶多肽（细胞趋化因子如CXCL11和肽类激素如GLP-1）N端的X-Ala or X-Pro基. CD26是一种高度保守、分布于多种细胞的，具有跨膜域和很短的胞质域，但常被剪切和释放作为可溶性环化4型，常形成自身二聚体或与FAP形成异二聚体。DPPⅣ的天然底物多种多样，在体内执行多种重要功能。它还是细胞膜受体和共刺激分子，从而参与机体的免疫调节、细胞移行、细胞黏附和细胞调亡过程，故与多种疾病的发病有关。各种DPP-IV抑制剂对II型糖尿病、自身免疫性疾病及细胞增殖性疾病（肿瘤）的治疗具有重要意义。DPP-4是重要的药物开发靶点，其中DPP-IV抑制剂Januvia磷酸西他列汀Sitagliptin  phosphate已作为抗糖尿病药物上市。