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timp1  (R&D Systems)


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    R&D Systems timp1
    Timp1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+timp1/Recombinant+Human+TIMP-1+Protein%2C+CF/pmc12065878-379-153-154
    Average 93 stars, based on 46 article reviews
    timp1 - by Bioz Stars, 2026-09
    93/100 stars

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    Purification:

    Article Title: The C-terminal domains of ADAMTS1 contain exosites involved in its proteoglycanase activity.
    Article Snippet: .. Semiquantitative proteoglycan cleavage assays Purified V1-5GAG (100 nM) was digested with ADAMTS1, in TNC-B buffer at 37 C for 2 h. Where indicated, 500 μM recombinant human TIMP1, TIMP2, TIMP3, or TIMP4 (R&D Systems, Cat. n.: 970-TM, 971-TM, 973-TM, 974-TSF) were preincubated with 100 nM ADAMTS1 for 1 h at 37 C before digestion. .. Biglycan (1130 nM) was digested for 24 h at 37 C. The reactions were stopped with EDTA (25 mM) in deglycosylation buffer (50 mM sodium acetate, 25 mM Tris–HCl pH 8.0) containing 0.1 U/ml chondroitinase ABC (AMS biotechnology Cat. n: AMS.E1028-02) for 16 h at 37 oC.

    Article Title: The C-terminal domains of ADAMTS1 contain exosites involved in its proteoglycanase activity
    Article Snippet: .. Purified V1-5GAG (100 nM) was digested with ADAMTS1, in TNC-B buffer at 37 °C for 2 h. Where indicated, 500 μM recombinant human TIMP1, TIMP2, TIMP3, or TIMP4 (R&D Systems, Cat. n.: 970-TM, 971-TM, 973-TM, 974-TSF) were preincubated with 100 nM ADAMTS1 for 1 h at 37 °C before digestion. ..

    Recombinant:

    Article Title: The C-terminal domains of ADAMTS1 contain exosites involved in its proteoglycanase activity.
    Article Snippet: .. Semiquantitative proteoglycan cleavage assays Purified V1-5GAG (100 nM) was digested with ADAMTS1, in TNC-B buffer at 37 C for 2 h. Where indicated, 500 μM recombinant human TIMP1, TIMP2, TIMP3, or TIMP4 (R&D Systems, Cat. n.: 970-TM, 971-TM, 973-TM, 974-TSF) were preincubated with 100 nM ADAMTS1 for 1 h at 37 C before digestion. .. Biglycan (1130 nM) was digested for 24 h at 37 C. The reactions were stopped with EDTA (25 mM) in deglycosylation buffer (50 mM sodium acetate, 25 mM Tris–HCl pH 8.0) containing 0.1 U/ml chondroitinase ABC (AMS biotechnology Cat. n: AMS.E1028-02) for 16 h at 37 oC.

    Article Title: Proteomic discovery of substrates of the cardiovascular protease ADAMTS7
    Article Snippet: .. Where TIMPs were used in the assay, recombinant human TIMP1, 2, 3 or 4 (R&D systems) were preincubated with ADAMTS7-T8 for 1 hour at 37° C. Proteolysis was stopped by addition of BoltTM LDS Sample Buffer, 5% β-mercaptoethanol and heating to 95° C. Samples were frozen at -20° C until analysis by SDS-PAGE/Coomassie. .. To measure TIMP-4 inhibition, proteolysis was quantified by densitometry using ImageJ.

    Article Title: LOXL2-mediated matrix remodeling in metastasis and mammary gland involution
    Article Snippet: Cells were transfected with SMARTpool siRNAs specific for human LOXL2(Dharmacon). .. Recombinant human TIMP1(R&D systems) was added to wells at a final concentration of 10nM. .. Immunohistochemistry/Immunofluorescence Staining was performed as previously described( 21 , 24 ) using antibodies described above plus cytokeratin 14(Abcam).

    Article Title: The C-terminal domains of ADAMTS1 contain exosites involved in its proteoglycanase activity
    Article Snippet: .. Purified V1-5GAG (100 nM) was digested with ADAMTS1, in TNC-B buffer at 37 °C for 2 h. Where indicated, 500 μM recombinant human TIMP1, TIMP2, TIMP3, or TIMP4 (R&D Systems, Cat. n.: 970-TM, 971-TM, 973-TM, 974-TSF) were preincubated with 100 nM ADAMTS1 for 1 h at 37 °C before digestion. ..

    Article Title: Loss of mutually protective effects between osteoclasts and chondrocytes in damaged joints drives osteoclast-mediated cartilage degradation via matrix metalloproteinases
    Article Snippet: .. For inhibitor experiments, mature osteoclasts were treated with bafilomycin (Cell Signalling Technology, Hitchin, UK), E64 (Cambridge Bioscience, Cambridge, UK), GM6001 (Selleckchem, Munich, Germany) or recombinant human TIMP1 (R&D Systems). .. Dentine (elephant ivory discs; HM Revenue & Customs, Heathrow Airport, UK) was prepared by cutting 250 μm transverse wafers using a low-speed saw and diamond-edged blade (Buehler, Coventry, UK), out of which 4 mm diameter discs were punched.

    Article Title: Loss of mutual protection between human osteoclasts and chondrocytes in damaged joints initiates osteoclast-mediated cartilage degradation by MMPs
    Article Snippet: .. For inhibitor experiments, mature osteoclasts were treated with bafilomycin (Cell Signalling Technology, Hitchin, UK), E64 (Cambridge Bioscience, Cambridge, UK), GM6001 (Selleckchem, Munich, Germany) or recombinant human TIMP1 (R&D Systems). .. Dentine (elephant ivory; HM Revenue & Customs, Heathrow Airport, UK) was prepared by cutting 250 μm transverse wafers using a low-speed saw and diamond-edged blade (Buehler, Coventry, UK) then punching out 4 mm diameter discs.

    Article Title: Proteomic discovery of substrates of the cardiovascular protease ADAMTS7
    Article Snippet: .. Where TIMPs were used in the assay, recombinant human TIMP1, 2, 3, or 4 (R&D Systems) were pre-incubated with ADAMTS7-T8 for 1 h at 37 °C. ..

    Concentration Assay:

    Article Title: LOXL2-mediated matrix remodeling in metastasis and mammary gland involution
    Article Snippet: Cells were transfected with SMARTpool siRNAs specific for human LOXL2(Dharmacon). .. Recombinant human TIMP1(R&D systems) was added to wells at a final concentration of 10nM. .. Immunohistochemistry/Immunofluorescence Staining was performed as previously described( 21 , 24 ) using antibodies described above plus cytokeratin 14(Abcam).



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    Figure 2. Inhibition of ADAMTS1 by TIMPs. A, inhibition of versicanase activity by TIMP family members. <t>TIMP1,</t> TIMP2, TIMP3, and TIMP4 (each at 500 nM) were incubated with ADAMTS1 (100 nM) for 1 h at 37 C before addition of V1-5GAG and digestion for 2 h. Following SDS-PAGE under reducing conditions (5% β-mercaptoethanol) and immunoblotting, FL V1-5GAG and versikine (VSK) were detected by the anti-Vc antibody. A representative immunoblot is shown (n = 2 independent experiments). B, inhibition of peptidolytic activity. TIMPs (each 25 nM) were incubated with a nominal con- centration of 25 nM ADAMTS1 for 1 h at 37 C before addition of the QF peptide substrate fluorescein-5(6)-carbonyl-Ala-Glu-Leu-Asn-Gly-Arg-Pro-Ile-Ser-Ile- Ala-Lys (5(6)-TAMRA) (3.5 μM) and digestion for 2 h. Following subtraction of the background (reactions not containing ADAMTS1), values were converted into percentage of ADAMTS1 activity in the absence of TIMPs and reported as average ± SD (n = 3, each point representing a technical replicate), p < 0.05 by Mann-Whitney test. C, titration of ADAMTS1 with TIMP3. TIMP3 (0–16 nM) was incubated with ADAMTS1 (20 nM nominal concentration) at 37 C for 1 h, and residual activity against the QF peptide FAM-AELNGRPISIAK-Tamra (3.5 μM) was determined. A representative titration curve is shown, each point representing a mean of two technical replicates. Final concentration of ADAMTS1 following titration was 10 nM. FL, full-length; No E, no enzyme; No I, no inhibitor; QF, Quenched-Fluorescent; TIMP, tissue inhibitor of metalloproteinase.
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    Image Search Results


    Figure 2. Inhibition of ADAMTS1 by TIMPs. A, inhibition of versicanase activity by TIMP family members. TIMP1, TIMP2, TIMP3, and TIMP4 (each at 500 nM) were incubated with ADAMTS1 (100 nM) for 1 h at 37 C before addition of V1-5GAG and digestion for 2 h. Following SDS-PAGE under reducing conditions (5% β-mercaptoethanol) and immunoblotting, FL V1-5GAG and versikine (VSK) were detected by the anti-Vc antibody. A representative immunoblot is shown (n = 2 independent experiments). B, inhibition of peptidolytic activity. TIMPs (each 25 nM) were incubated with a nominal con- centration of 25 nM ADAMTS1 for 1 h at 37 C before addition of the QF peptide substrate fluorescein-5(6)-carbonyl-Ala-Glu-Leu-Asn-Gly-Arg-Pro-Ile-Ser-Ile- Ala-Lys (5(6)-TAMRA) (3.5 μM) and digestion for 2 h. Following subtraction of the background (reactions not containing ADAMTS1), values were converted into percentage of ADAMTS1 activity in the absence of TIMPs and reported as average ± SD (n = 3, each point representing a technical replicate), p < 0.05 by Mann-Whitney test. C, titration of ADAMTS1 with TIMP3. TIMP3 (0–16 nM) was incubated with ADAMTS1 (20 nM nominal concentration) at 37 C for 1 h, and residual activity against the QF peptide FAM-AELNGRPISIAK-Tamra (3.5 μM) was determined. A representative titration curve is shown, each point representing a mean of two technical replicates. Final concentration of ADAMTS1 following titration was 10 nM. FL, full-length; No E, no enzyme; No I, no inhibitor; QF, Quenched-Fluorescent; TIMP, tissue inhibitor of metalloproteinase.

    Journal: The Journal of biological chemistry

    Article Title: The C-terminal domains of ADAMTS1 contain exosites involved in its proteoglycanase activity.

    doi: 10.1016/j.jbc.2023.103048

    Figure Lengend Snippet: Figure 2. Inhibition of ADAMTS1 by TIMPs. A, inhibition of versicanase activity by TIMP family members. TIMP1, TIMP2, TIMP3, and TIMP4 (each at 500 nM) were incubated with ADAMTS1 (100 nM) for 1 h at 37 C before addition of V1-5GAG and digestion for 2 h. Following SDS-PAGE under reducing conditions (5% β-mercaptoethanol) and immunoblotting, FL V1-5GAG and versikine (VSK) were detected by the anti-Vc antibody. A representative immunoblot is shown (n = 2 independent experiments). B, inhibition of peptidolytic activity. TIMPs (each 25 nM) were incubated with a nominal con- centration of 25 nM ADAMTS1 for 1 h at 37 C before addition of the QF peptide substrate fluorescein-5(6)-carbonyl-Ala-Glu-Leu-Asn-Gly-Arg-Pro-Ile-Ser-Ile- Ala-Lys (5(6)-TAMRA) (3.5 μM) and digestion for 2 h. Following subtraction of the background (reactions not containing ADAMTS1), values were converted into percentage of ADAMTS1 activity in the absence of TIMPs and reported as average ± SD (n = 3, each point representing a technical replicate), p < 0.05 by Mann-Whitney test. C, titration of ADAMTS1 with TIMP3. TIMP3 (0–16 nM) was incubated with ADAMTS1 (20 nM nominal concentration) at 37 C for 1 h, and residual activity against the QF peptide FAM-AELNGRPISIAK-Tamra (3.5 μM) was determined. A representative titration curve is shown, each point representing a mean of two technical replicates. Final concentration of ADAMTS1 following titration was 10 nM. FL, full-length; No E, no enzyme; No I, no inhibitor; QF, Quenched-Fluorescent; TIMP, tissue inhibitor of metalloproteinase.

    Article Snippet: Semiquantitative proteoglycan cleavage assays Purified V1-5GAG (100 nM) was digested with ADAMTS1, in TNC-B buffer at 37 C for 2 h. Where indicated, 500 μM recombinant human TIMP1, TIMP2, TIMP3, or TIMP4 (R&D Systems, Cat. n.: 970-TM, 971-TM, 973-TM, 974-TSF) were preincubated with 100 nM ADAMTS1 for 1 h at 37 C before digestion.

    Techniques: Inhibition, Activity Assay, Incubation, SDS Page, Western Blot, MANN-WHITNEY, Titration, Concentration Assay