推薦產品
生物源
human
品質等級
形狀
liquid
製造商／商標名
Chemicon®
濃度
>50% (truncated ADAMTS-5 of total protein of the preparation, SDS-PAGE)
0.1 mg/mL (total protein concentration)
0.1 mg/mL
NCBI登錄號
UniProt登錄號
運輸包裝
dry ice
基因資訊
human ... ADAMTS5(11096)
一般說明
ADAMTS′s (a disintegrin and metalloproteinase with thrombospondin motif) are multidomain extracellular proteinases, containing at least one thrombospondin type-1 repeat (Tang, 2001). The enzymes are involved in processing of extracellular matrix proteins and proteins in the circulation. ADAMTS-5 was first purified from bovine nasal cartilage conditioned media and human ADAMTS-5 cDNA was cloned from a human liver cDNA library (Abbaszade, I. et al, 1999). ADAMTS-5 consists of a prodomain, a catalytic domain, a disintegrin domain, a thrombospondin type-1 motif, a Cys-rich region followed by a spacer sequence and thrombospondin submotifs. The prodomain is most likely cleaved-off by a furin-type enzyme before ADAMTS-5 is released from cells. ADAMTS-5 is expressed constitutively in synovium (Vankemmelbeke, MN. et al, 2001) and it degrades aggrecan, the major proteoglycan of articular cartilage. Like ADAMTS4, another member of the ADAMTS-family, ADAMTS-5 cleaves aggrecan at 5 sites (Tortorella, MD. et al, 2002). Four cleavage sites are located in the chondroitin sulfate-rich region between aggrecan globular domains G2 and G3, while one site is contained in the rod-shaped polypeptide between globular domains G1 and G2. In addition, ADAMTS-5 cleaves one more site in the chondroitin sulfate-rich region that is not cleaved by ADAMTS4 (Tortorella, MD. et al, 2002). ADAMTS-5 is inhibited by TIMP 3 (Kashigawa, M. et al, 2001) and a2-macroglobulin (Tortorella, MD. et al, 2004).
Molecular form: Recombinant human ADAMTS-5 D625-930 is produced with the baculo-virus expression system and purified from insect cell culture supernatants. The protein contains the catalytic domain, the disintegrin domain and the thrombospondin type-1 motif of full-length ADAMTS-5 followed by a C-terminal His6-tag. The calculated Mr of the amino acid sequence is 40.8 kDa. In SDS-PAGE the protein exhibits a Mr of about 50 kDa.
ACTIVITY:
Aggrecanase activity of the ADAMTS-5 preparation is determined with recombinant aggrecan interglobular domain (Aggrecan-IGD from Chemicon). ADAMTS-5 hydrolyzes the "aggrecanase" site within this domain (peptide bond E373 -A374 in human aggrecan). The recombinant substrate is incubated at a concentration of 0.1 μM with ADAMTS-5 in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 1 μM leupeptin, 1 μM pepstatin, 1 mM Pefabloc, 0.05 % Brij 35 for 15 min at 37 °C. Substrate cleavage at the "aggrecanase"-site is estimated from the appearance of the hydrolysis fragment with the novel N-terminus ARGSVIL. The fragment is quantified with two monoclonal antibodies. Under the specified conditions the hydrolysis rate is >0.5 nmoles hydrolyzed substrate/ min . ml ADAMTS-5 preparation or >5 nmoles hydrolyzed substrate/ min . mg.
Inhibitors: ADAMTS-5 is inhibited by tissue inhibitor of matrix metalloproteinase 3 (TIMP3) and by alpha2-macroglobulin. Enzyme activity is also suppressed by chelators of divalent cations such as EDTA and by synthetic metalloproteinase inhibitors.
Molecular form: Recombinant human ADAMTS-5 D625-930 is produced with the baculo-virus expression system and purified from insect cell culture supernatants. The protein contains the catalytic domain, the disintegrin domain and the thrombospondin type-1 motif of full-length ADAMTS-5 followed by a C-terminal His6-tag. The calculated Mr of the amino acid sequence is 40.8 kDa. In SDS-PAGE the protein exhibits a Mr of about 50 kDa.
ACTIVITY:
Aggrecanase activity of the ADAMTS-5 preparation is determined with recombinant aggrecan interglobular domain (Aggrecan-IGD from Chemicon). ADAMTS-5 hydrolyzes the "aggrecanase" site within this domain (peptide bond E373 -A374 in human aggrecan). The recombinant substrate is incubated at a concentration of 0.1 μM with ADAMTS-5 in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 1 μM leupeptin, 1 μM pepstatin, 1 mM Pefabloc, 0.05 % Brij 35 for 15 min at 37 °C. Substrate cleavage at the "aggrecanase"-site is estimated from the appearance of the hydrolysis fragment with the novel N-terminus ARGSVIL. The fragment is quantified with two monoclonal antibodies. Under the specified conditions the hydrolysis rate is >0.5 nmoles hydrolyzed substrate/ min . ml ADAMTS-5 preparation or >5 nmoles hydrolyzed substrate/ min . mg.
Inhibitors: ADAMTS-5 is inhibited by tissue inhibitor of matrix metalloproteinase 3 (TIMP3) and by alpha2-macroglobulin. Enzyme activity is also suppressed by chelators of divalent cations such as EDTA and by synthetic metalloproteinase inhibitors.
應用
Functional Studies
Degradation of extracellular matrix proteoglycans
Screening and characterisation of inhibitors
Standard in enzymatic and immunological assays
Optimal working dilutions must be determined by the end user.
Degradation of extracellular matrix proteoglycans
Screening and characterisation of inhibitors
Standard in enzymatic and immunological assays
Optimal working dilutions must be determined by the end user.
外觀
Solubilized in 50mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 50mM imidazol 0.05 % Brij-35.
儲存和穩定性
Maintain at -70°C for 12 months from date of receipt. The enzyme can be kept at -20°C for several weeks and ice for several days. Avoid repeated freeze/thaw cycles.
法律資訊
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
訊號詞
Danger
危險聲明
危險分類
Repr. 1B
儲存類別代碼
6.1D - Non-combustible acute toxic Cat.3 / toxic hazardous materials or hazardous materials causing chronic effects
水污染物質分類（WGK）
WGK 1
閃點（°F）
Not applicable
閃點（°C）
Not applicable
分析證明 (COA)
輸入產品批次/批號來搜索 分析證明 (COA)。在產品’s標籤上找到批次和批號，寫有 ‘Lot’或‘Batch’.。
M N Vankemmelbeke et al.
European journal of biochemistry, 268(5), 1259-1268 (2001-03-07)
ADAMTS proteinases, belonging to the adamalysin subfamily of metalloproteinases, have been implicated in a variety of cellular events such as morphogenesis, cell migration, angiogenesis, ovulation and extracellular matrix breakdown. Aggrecanase-1 (ADAMTS-4) and aggrecanase-2 (ADAMTS-5) have been identified in cartilage and
M Kashiwagi et al.
The Journal of biological chemistry, 276(16), 12501-12504 (2001-03-30)
The proteoglycan aggrecan is an important major component of cartilage matrix that gives articular cartilage the ability to withstand compression. Increased breakdown of aggrecan is associated with the development of arthritis and is considered to be catalyzed by aggrecanases, members
Micky D Tortorella et al.
Matrix biology : journal of the International Society for Matrix Biology, 21(6), 499-511 (2002-10-24)
ADAM-TS5 (aggrecanase 2), one of two cartilage aggrecanases is a member of the ADAM protein family. Like ADAM-TS4 (aggrecanase 1) the enzyme cleaves cartilage aggrecan at the Glu(373)-Ala(374) bond, a marker of aggrecanase activity. In this study we have characterized
B L Tang
The international journal of biochemistry & cell biology, 33(1), 33-44 (2001-02-13)
ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) is a novel family of extracellular proteases found in both mammals and invertebrates. Members of the family may be distinguished from the ADAM (a disintegrin and metalloprotease) family members based on the
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