recombinant timp 1  (New England Biolabs)


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  • 88
    Name:
    alpha 1 2 Fucosidase
    Description:
    alpha 1 2 Fucosidase 5 000 units
    Catalog Number:
    p0724l
    Price:
    520
    Size:
    5 000 units
    Category:
    Glycosidases
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    Structured Review

    New England Biolabs recombinant timp 1
    alpha 1 2 Fucosidase
    alpha 1 2 Fucosidase 5 000 units
    https://www.bioz.com/result/recombinant timp 1/product/New England Biolabs
    Average 88 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    recombinant timp 1 - by Bioz Stars, 2020-07
    88/100 stars

    Images

    1) Product Images from "Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)"

    Article Title: Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)

    Journal: Clinical proteomics

    doi: 10.1186/1559-0275-10-12

    Schematic representation of the dLISA approach. (A) A serum sample is split into five aliquots (A1 to A5). (B) Four of these aliquots are treated with increasing amounts of magnetic beads coupled with TIMP-1 antibody to gradually reduce the amount of endogenous TIMP-1 present. For aliquot A5, there should be sufficient beads used for complete removal of the endogenous TIMP-1. (C) After the magnetic beads treatment, the beads were separated using a magnetic separator and the supernatant from each aliquot is transferred. (D) Both TIMP-1 UEA LISA assay and TIMP-1 immunoassay are performed on each aliquot. (E) A dose–response curve is constructed using the measured TIMP-1 concentration (ng/mL) in each aliquot by the TIMP-1 immunoassay along with the corresponding fluorescence readout by the TIMP-1 UEA LISA assay.
    Figure Legend Snippet: Schematic representation of the dLISA approach. (A) A serum sample is split into five aliquots (A1 to A5). (B) Four of these aliquots are treated with increasing amounts of magnetic beads coupled with TIMP-1 antibody to gradually reduce the amount of endogenous TIMP-1 present. For aliquot A5, there should be sufficient beads used for complete removal of the endogenous TIMP-1. (C) After the magnetic beads treatment, the beads were separated using a magnetic separator and the supernatant from each aliquot is transferred. (D) Both TIMP-1 UEA LISA assay and TIMP-1 immunoassay are performed on each aliquot. (E) A dose–response curve is constructed using the measured TIMP-1 concentration (ng/mL) in each aliquot by the TIMP-1 immunoassay along with the corresponding fluorescence readout by the TIMP-1 UEA LISA assay.

    Techniques Used: Magnetic Beads, Construct, Concentration Assay, Fluorescence

    Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).
    Figure Legend Snippet: Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).

    Techniques Used: Recombinant

    Dose-response curves and slopes of the dLISA approach when applied to serum specimens with the same fucosylated TIMP-1 in different serum matrices. (A) Dose–response curves of four samples with the same fucosylated TIMP1 in different matrices. For preparation of these specimens, we identified four individual serum samples. Each one of them was then treated for complete removal of endogenous TIMP-1 (confirmed by the TIMP-1 immunoassay). Recombinant TIMP-1 was added into each of the serum matrices. Then these serum samples underwent the dLISA approach. (B) A bar graph with linear regression slopes of the dose–response curves.
    Figure Legend Snippet: Dose-response curves and slopes of the dLISA approach when applied to serum specimens with the same fucosylated TIMP-1 in different serum matrices. (A) Dose–response curves of four samples with the same fucosylated TIMP1 in different matrices. For preparation of these specimens, we identified four individual serum samples. Each one of them was then treated for complete removal of endogenous TIMP-1 (confirmed by the TIMP-1 immunoassay). Recombinant TIMP-1 was added into each of the serum matrices. Then these serum samples underwent the dLISA approach. (B) A bar graph with linear regression slopes of the dose–response curves.

    Techniques Used: Recombinant

    2) Product Images from "Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)"

    Article Title: Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)

    Journal: Clinical proteomics

    doi: 10.1186/1559-0275-10-12

    Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).
    Figure Legend Snippet: Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).

    Techniques Used: Recombinant

    3) Product Images from "Human stem cell-derived retinal epithelial cells activate complement via collectin 11 in response to stress"

    Article Title: Human stem cell-derived retinal epithelial cells activate complement via collectin 11 in response to stress

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-15212-z

    C3d expression and MAC formation on hypoxia-stressed iPS-RPE cells following fucosidase treatment. ( a ) Representative immunofluorescence images showing C3d deposition on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. C3d (green), nuclei nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm. ( b ) Quantification of C3d expression level on hypoxia-stressed iPS- RPE cells compared to fucosidase-treated hypoxic cells using ImageJ software. ( c ) Representative immunofluorescence images showing MAC staining on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. MAC (red), nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm.
    Figure Legend Snippet: C3d expression and MAC formation on hypoxia-stressed iPS-RPE cells following fucosidase treatment. ( a ) Representative immunofluorescence images showing C3d deposition on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. C3d (green), nuclei nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm. ( b ) Quantification of C3d expression level on hypoxia-stressed iPS- RPE cells compared to fucosidase-treated hypoxic cells using ImageJ software. ( c ) Representative immunofluorescence images showing MAC staining on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. MAC (red), nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm.

    Techniques Used: Expressing, Immunofluorescence, Software, Staining

    4) Product Images from "Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)"

    Article Title: Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)

    Journal: Clinical proteomics

    doi: 10.1186/1559-0275-10-12

    Schematic representation of the dLISA approach. (A) A serum sample is split into five aliquots (A1 to A5). (B) Four of these aliquots are treated with increasing amounts of magnetic beads coupled with TIMP-1 antibody to gradually reduce the amount of endogenous TIMP-1 present. For aliquot A5, there should be sufficient beads used for complete removal of the endogenous TIMP-1. (C) After the magnetic beads treatment, the beads were separated using a magnetic separator and the supernatant from each aliquot is transferred. (D) Both TIMP-1 UEA LISA assay and TIMP-1 immunoassay are performed on each aliquot. (E) A dose–response curve is constructed using the measured TIMP-1 concentration (ng/mL) in each aliquot by the TIMP-1 immunoassay along with the corresponding fluorescence readout by the TIMP-1 UEA LISA assay.
    Figure Legend Snippet: Schematic representation of the dLISA approach. (A) A serum sample is split into five aliquots (A1 to A5). (B) Four of these aliquots are treated with increasing amounts of magnetic beads coupled with TIMP-1 antibody to gradually reduce the amount of endogenous TIMP-1 present. For aliquot A5, there should be sufficient beads used for complete removal of the endogenous TIMP-1. (C) After the magnetic beads treatment, the beads were separated using a magnetic separator and the supernatant from each aliquot is transferred. (D) Both TIMP-1 UEA LISA assay and TIMP-1 immunoassay are performed on each aliquot. (E) A dose–response curve is constructed using the measured TIMP-1 concentration (ng/mL) in each aliquot by the TIMP-1 immunoassay along with the corresponding fluorescence readout by the TIMP-1 UEA LISA assay.

    Techniques Used: Magnetic Beads, Construct, Concentration Assay, Fluorescence

    Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).
    Figure Legend Snippet: Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).

    Techniques Used: Recombinant

    Dose-response curves and slopes of the dLISA approach when applied to serum specimens with the same fucosylated TIMP-1 in different serum matrices. (A) Dose–response curves of four samples with the same fucosylated TIMP1 in different matrices. For preparation of these specimens, we identified four individual serum samples. Each one of them was then treated for complete removal of endogenous TIMP-1 (confirmed by the TIMP-1 immunoassay). Recombinant TIMP-1 was added into each of the serum matrices. Then these serum samples underwent the dLISA approach. (B) A bar graph with linear regression slopes of the dose–response curves.
    Figure Legend Snippet: Dose-response curves and slopes of the dLISA approach when applied to serum specimens with the same fucosylated TIMP-1 in different serum matrices. (A) Dose–response curves of four samples with the same fucosylated TIMP1 in different matrices. For preparation of these specimens, we identified four individual serum samples. Each one of them was then treated for complete removal of endogenous TIMP-1 (confirmed by the TIMP-1 immunoassay). Recombinant TIMP-1 was added into each of the serum matrices. Then these serum samples underwent the dLISA approach. (B) A bar graph with linear regression slopes of the dose–response curves.

    Techniques Used: Recombinant

    Related Articles

    Incubation:

    Article Title: Human Norovirus Histo-Blood Group Antigen (HBGA) Binding Sites Mediate the Virus Specific Interactions with Lettuce Carbohydrates
    Article Snippet: .. For fucosidase digestion, each slide was digested with 20 U α-1,2 fucosidase (NEB, Ipswich, MA, USA) at 37 °C for 1 h. For inhibition with HBGA monoclonal antibody or lectins, a 200 µL of anti-H type 1 HBGA monoclonal antibody BG4 (Covance, Emeryville, CA, USA) diluted in PBS were added to the slides and incubated for 1 h at 37 °C. .. Plant lectins DBA (Dolichos biflorus ), LEA (Lycopersicon esculentum ), BS-I (Bandeiraea simplicifolia ) and LcH (Lens culinaris ), which recognize GalNAc (type A HBGA), GlcNAc, α-D-Gal (type B HBGA) and α-D-Man/α-D-Glc, respectively, were diluted in TBS buffer (50 mM Tris-HCl, 150 mM NaCl, supplemented with 2.5 mM MnCl2 and CaCl2 ) to a final concentration of 100 µg/mL added to the slides and incubated for 1 h at 37 °C.

    Recombinant:

    Article Title: Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)
    Article Snippet: .. Production of the differential fucosylated recombinant TIMP-1 Four microliter of the recombinant TIMP-1 (10 μg/mL prepared in PBS + 1% BSA buffer) was mixed with 4 μL of α1, 2 fucosidase (Catalog# P0724, New England Biolabs, Ipswich, MA). .. The mixture (8 μL) was then diluted 5 times in the 32 μL of the Reaction buffer (50 mM Sodium Citrate, 100 mM Sodium Chloride, pH 6.0) for incubation of 0, 15, 30, or 45 minutes with shaking at 37°C.

    Inhibition:

    Article Title: Human Norovirus Histo-Blood Group Antigen (HBGA) Binding Sites Mediate the Virus Specific Interactions with Lettuce Carbohydrates
    Article Snippet: .. For fucosidase digestion, each slide was digested with 20 U α-1,2 fucosidase (NEB, Ipswich, MA, USA) at 37 °C for 1 h. For inhibition with HBGA monoclonal antibody or lectins, a 200 µL of anti-H type 1 HBGA monoclonal antibody BG4 (Covance, Emeryville, CA, USA) diluted in PBS were added to the slides and incubated for 1 h at 37 °C. .. Plant lectins DBA (Dolichos biflorus ), LEA (Lycopersicon esculentum ), BS-I (Bandeiraea simplicifolia ) and LcH (Lens culinaris ), which recognize GalNAc (type A HBGA), GlcNAc, α-D-Gal (type B HBGA) and α-D-Man/α-D-Glc, respectively, were diluted in TBS buffer (50 mM Tris-HCl, 150 mM NaCl, supplemented with 2.5 mM MnCl2 and CaCl2 ) to a final concentration of 100 µg/mL added to the slides and incubated for 1 h at 37 °C.

    Blocking Assay:

    Article Title: Recognition of Histo-Blood Group Antigen-Like Carbohydrates in Lettuce by Human GII.4 Norovirus
    Article Snippet: .. For fucosidase digestion, each slide was digested with 20 U of α-1,2-fucosidase (NEB, Ipswich, MA) at 37°C for 1 h. Following three washes with 300 ml of phosphate-buffered saline (PBS) containing 0.05% Tween 20 (PBS-T), blocking was performed at RT with PBS supplemented with 1% bovine serum albumin (BSA). .. Subsequently, the slides were incubated with 200 μl of NoV VLPs (final concentration, 10 μg/ml) at RT for 1 h. The HuNoV VLPs on lettuce were detected by immunofluorescence assay (IFA) as follows.

    Article Title: Bacterial AB5 toxins inhibit the growth of gut bacteria by targeting ganglioside-like glycoconjugates
    Article Snippet: .. Where indicated, slides were treated with α1-2-fucosidase (NEB, P0724S) following antigen retrieval in sodium citrate buffer, but prior to blocking with donkey serum buffer. .. Slides were treated for 1 h, with 1 μl (20 units) of α1-2-fucosidase in 10 μl of 1× Glycobuffer (supplied by the manufacturer), at 37 °C, followed by a PBS wash. Each slide was then treated with CTB (1 mM, Sigma, Cat. No. C9903) for 1 h, followed by either α-CTB antibody (1:100) or α-GM1 antibody (1:100, Abcam) for 1 h. Slides were visualized using Alexa Fluor 488-conjugated donkey α-rabbit IgG (1:1000, Invitrogen).

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  • 94
    New England Biolabs fucosidase
    Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 <t>fucosidase</t> at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).
    Fucosidase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fucosidase/product/New England Biolabs
    Average 94 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    fucosidase - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).

    Journal: Clinical proteomics

    Article Title: Analysis of serum protein glycosylation by a differential lectin immunosorbant assay (dLISA)

    doi: 10.1186/1559-0275-10-12

    Figure Lengend Snippet: Slopes of the dose response curves established using the dLISA approach correlated with TIMP-1 fucosylation. (A) Dose–response curves of serum samples spiked with differentially fucosylated recombinant TIMP-1. Recombinant TIMP-1 was treated by α1, 2 fucosidase at different length of time (0, 15, 30, and 45 minutes) before they were spiked into with four serum aliquots. These aliquots came from the same pool of sera with endogenous TIMP-1 immuno-depleted (confirmed by the TIMP-1 immunoassay). After the recombinant protein spiked in, these serum samples underwent the dLISA approach. (B) A graph presentation of linear regression slopes of the dose–response curves versus the length of fucosidase treatment (minutes).

    Article Snippet: Production of the differential fucosylated recombinant TIMP-1 Four microliter of the recombinant TIMP-1 (10 μg/mL prepared in PBS + 1% BSA buffer) was mixed with 4 μL of α1, 2 fucosidase (Catalog# P0724, New England Biolabs, Ipswich, MA).

    Techniques: Recombinant

    C3d expression and MAC formation on hypoxia-stressed iPS-RPE cells following fucosidase treatment. ( a ) Representative immunofluorescence images showing C3d deposition on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. C3d (green), nuclei nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm. ( b ) Quantification of C3d expression level on hypoxia-stressed iPS- RPE cells compared to fucosidase-treated hypoxic cells using ImageJ software. ( c ) Representative immunofluorescence images showing MAC staining on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. MAC (red), nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm.

    Journal: Scientific Reports

    Article Title: Human stem cell-derived retinal epithelial cells activate complement via collectin 11 in response to stress

    doi: 10.1038/s41598-017-15212-z

    Figure Lengend Snippet: C3d expression and MAC formation on hypoxia-stressed iPS-RPE cells following fucosidase treatment. ( a ) Representative immunofluorescence images showing C3d deposition on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. C3d (green), nuclei nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm. ( b ) Quantification of C3d expression level on hypoxia-stressed iPS- RPE cells compared to fucosidase-treated hypoxic cells using ImageJ software. ( c ) Representative immunofluorescence images showing MAC staining on the surface of hypoxia-stressed iPS-RPE cells treated or not overnight with fucosidase. MAC (red), nuclei (blue) and phalloidin (grey) are shown. Scale bars, 50 μm.

    Article Snippet: To remove L-fucose from cell surface, iPS-RPE cells were put under hypoxic conditions for 24 hours, fixed in 4% paraformaldehyde for 1 hour at room temperature and treated then with α1-2,3,4,6 fucosidase (New England BioLabs) at 37 °C overnight.

    Techniques: Expressing, Immunofluorescence, Software, Staining