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Seikagaku flavobacterium heparinum heparitinase
Flavobacterium Heparinum Heparitinase, supplied by Seikagaku, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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flavobacterium heparinum heparitinase - by Bioz Stars, 2020-11
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Article Title: Impairment of Embryonic Cell Division and Glycosaminoglycan Biosynthesis in Glucuronyltransferase-I-deficient Mice *
Article Snippet: Proteus vulgaris chondroitinase ABC (CSase) (EC 4.2.2.4), Flavobacterium heparinum heparitinase and heparinase, and the monoclonal antibodies (LY111, Hepss-1, and 3G10) were purchased from Seikagaku Corp. (Tokyo, Japan).

Article Title: Chondroitin Sulfate Is Indispensable for Pluripotency and Differentiation of Mouse Embryonic Stem Cells
Article Snippet: Materials Proteus vulgaris chondroitinase ABC (CSase; EC 4.2.2.4), Flavobacterium heparinum heparitinase and heparinase, the monoclonal antibodies Hepss-1 and LY111, CS-A from whale cartilage, CS-C from shark cartilage, CS-E from squid cartilage, and HS from bovine kidneys were purchased from Seikagaku Corp. (Tokyo, Japan).

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    Seikagaku heparitinase i
    Western blotting analysis of DRM fractions isolated from a rat parathyroid cell line. DRMs were prepared from confluent PTr cells as described in Materials and Methods . Collected fractions, were concentrated, treated with <t>heparitinase</t> I and subjected to SDS-PAGE and WB analysis. A. Staining with anti-ΔHS (3G10) antibodies confirmed the presence of HSPGs in low-density fractions. Equal volumes (13 µl) of each fraction were analyzed. Fractions 13 and 14, bottom fraction (pooled fractions 15 and 16, B) and lysate (L) were diluted 16, 62 and 56 times, respectively, prior to the analysis. Bands marked with (*) represent non-specific staining due to the presence of BSA at high concentration. B. Staining with antibodies against DRM markers, Lyn and Giα defined the low-density fractions as DRMs. Equal volumes (33 µl) of each fraction were used for analysis. Fractions 13 through 14, bottom fraction (pooled fractions 15 and 16, B) and lysate (L) were diluted 18, 72 and 64 times, respectively, prior the analysis due to high protein content. Staining for transferrin receptor (TfR) was used as a control for the successful preparation. C. Graphic representation of the distribution of TfR, Lyn, Giα and HSPGs in fractions obtained from sucrose-density gradient ultracentrifugation. Density of bands detected in WB analysis (A and C) was measured and expressed as arbitrary units. TfR (○); Lyn (▪); Giα (◊) and HSPG (▴).
    Heparitinase I, supplied by Seikagaku, used in various techniques. Bioz Stars score: 91/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/heparitinase i/product/Seikagaku
    Average 91 stars, based on 45 article reviews
    Price from $9.99 to $1999.99
    heparitinase i - by Bioz Stars, 2020-11
    91/100 stars
      Buy from Supplier

    85
    Seikagaku flavobacterium heparitinase
    Localization of DS and HS in uninjured carotid arteries of wild-type mice. Frozen sections were treated with chondroitin B-lyase (A), Flavobacterium <t>heparitinase</t> (C), or buffer alone (B,D) and then incubated with monoclonal antibodies ΔDi-4S (A,B) or ΔHS (C,D). Bound monoclonal antibodies were detected with a peroxidase-conjugated secondary antibody. Arrow indicates internal elastic lamina; arrowhead, external elastic lamina. DS was present primarily in the adventitia (A) and HS in the intima/media (C).
    Flavobacterium Heparitinase, supplied by Seikagaku, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flavobacterium heparitinase/product/Seikagaku
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    flavobacterium heparitinase - by Bioz Stars, 2020-11
    85/100 stars
      Buy from Supplier

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    Western blotting analysis of DRM fractions isolated from a rat parathyroid cell line. DRMs were prepared from confluent PTr cells as described in Materials and Methods . Collected fractions, were concentrated, treated with heparitinase I and subjected to SDS-PAGE and WB analysis. A. Staining with anti-ΔHS (3G10) antibodies confirmed the presence of HSPGs in low-density fractions. Equal volumes (13 µl) of each fraction were analyzed. Fractions 13 and 14, bottom fraction (pooled fractions 15 and 16, B) and lysate (L) were diluted 16, 62 and 56 times, respectively, prior to the analysis. Bands marked with (*) represent non-specific staining due to the presence of BSA at high concentration. B. Staining with antibodies against DRM markers, Lyn and Giα defined the low-density fractions as DRMs. Equal volumes (33 µl) of each fraction were used for analysis. Fractions 13 through 14, bottom fraction (pooled fractions 15 and 16, B) and lysate (L) were diluted 18, 72 and 64 times, respectively, prior the analysis due to high protein content. Staining for transferrin receptor (TfR) was used as a control for the successful preparation. C. Graphic representation of the distribution of TfR, Lyn, Giα and HSPGs in fractions obtained from sucrose-density gradient ultracentrifugation. Density of bands detected in WB analysis (A and C) was measured and expressed as arbitrary units. TfR (○); Lyn (▪); Giα (◊) and HSPG (▴).

    Journal: PLoS ONE

    Article Title: Syndecans Reside in Sphingomyelin-Enriched Low-Density Fractions of the Plasma Membrane Isolated from a Parathyroid Cell Line

    doi: 10.1371/journal.pone.0032351

    Figure Lengend Snippet: Western blotting analysis of DRM fractions isolated from a rat parathyroid cell line. DRMs were prepared from confluent PTr cells as described in Materials and Methods . Collected fractions, were concentrated, treated with heparitinase I and subjected to SDS-PAGE and WB analysis. A. Staining with anti-ΔHS (3G10) antibodies confirmed the presence of HSPGs in low-density fractions. Equal volumes (13 µl) of each fraction were analyzed. Fractions 13 and 14, bottom fraction (pooled fractions 15 and 16, B) and lysate (L) were diluted 16, 62 and 56 times, respectively, prior to the analysis. Bands marked with (*) represent non-specific staining due to the presence of BSA at high concentration. B. Staining with antibodies against DRM markers, Lyn and Giα defined the low-density fractions as DRMs. Equal volumes (33 µl) of each fraction were used for analysis. Fractions 13 through 14, bottom fraction (pooled fractions 15 and 16, B) and lysate (L) were diluted 18, 72 and 64 times, respectively, prior the analysis due to high protein content. Staining for transferrin receptor (TfR) was used as a control for the successful preparation. C. Graphic representation of the distribution of TfR, Lyn, Giα and HSPGs in fractions obtained from sucrose-density gradient ultracentrifugation. Density of bands detected in WB analysis (A and C) was measured and expressed as arbitrary units. TfR (○); Lyn (▪); Giα (◊) and HSPG (▴).

    Article Snippet: Biotinylated mouse anti-ΔHS (3G10) antibodies, recognizing HS neo-epitope, generated by the digestion with heparitinase I from Flavobacterium heparinum and heparitinase I (Flavobacterium heparinum ) were purchased from Seikagaku Corporation, (Tokyo, Japan).

    Techniques: Western Blot, Isolation, SDS Page, Staining, Concentration Assay

    Identification of HSPGs expressed by a rat parathyroid cell line. A. RT-PCR analysis of PTr cells using syndecan-specific primers (see Materials and Methods for details). Total RNA was isolated from confluent cells and subjected to RT-PCR analysis. Amplified products were run on 2% agarose gel, stained with ethidium bromide and photographed under UV transilluminator. Lanes: M – 100 bp marker; SN1 – amplification with syndecan-1 specific primers; SN2 – amplification with syndecan-2 specific primers; SN3 – amplification with syndecan-3 specific primers; SN4 – amplification with syndecan-4 specific primers; G – amplification with GAPHD specific primers; (-) – negative controls containing no cDNA. B. Identification of HSPGs present in DRM fractions using WB analysis. Proteoglycans were isolated from confluent rat parathyroid cells and partially purified using Q-Sepharose anion-exchange chromatography. A proteoglycan-enriched fraction was incubated in the presence or absence of heparitinase I, subjected to SDS-PAGE and immunoblotted with anti-syndecan-1, anti-syndecan-4 or anti-ΔHS (3G10) antibodies. Lanes: 1, 4 and 7 represent the heparitinase I only; 2, 5 and 8 correspond to the control samples, incubated without heparitinase I; 3, 6, 8 correspond to the heparitinase-treated samples.

    Journal: PLoS ONE

    Article Title: Syndecans Reside in Sphingomyelin-Enriched Low-Density Fractions of the Plasma Membrane Isolated from a Parathyroid Cell Line

    doi: 10.1371/journal.pone.0032351

    Figure Lengend Snippet: Identification of HSPGs expressed by a rat parathyroid cell line. A. RT-PCR analysis of PTr cells using syndecan-specific primers (see Materials and Methods for details). Total RNA was isolated from confluent cells and subjected to RT-PCR analysis. Amplified products were run on 2% agarose gel, stained with ethidium bromide and photographed under UV transilluminator. Lanes: M – 100 bp marker; SN1 – amplification with syndecan-1 specific primers; SN2 – amplification with syndecan-2 specific primers; SN3 – amplification with syndecan-3 specific primers; SN4 – amplification with syndecan-4 specific primers; G – amplification with GAPHD specific primers; (-) – negative controls containing no cDNA. B. Identification of HSPGs present in DRM fractions using WB analysis. Proteoglycans were isolated from confluent rat parathyroid cells and partially purified using Q-Sepharose anion-exchange chromatography. A proteoglycan-enriched fraction was incubated in the presence or absence of heparitinase I, subjected to SDS-PAGE and immunoblotted with anti-syndecan-1, anti-syndecan-4 or anti-ΔHS (3G10) antibodies. Lanes: 1, 4 and 7 represent the heparitinase I only; 2, 5 and 8 correspond to the control samples, incubated without heparitinase I; 3, 6, 8 correspond to the heparitinase-treated samples.

    Article Snippet: Biotinylated mouse anti-ΔHS (3G10) antibodies, recognizing HS neo-epitope, generated by the digestion with heparitinase I from Flavobacterium heparinum and heparitinase I (Flavobacterium heparinum ) were purchased from Seikagaku Corporation, (Tokyo, Japan).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Agarose Gel Electrophoresis, Staining, Marker, Western Blot, Purification, Chromatography, Incubation, SDS Page

    Treatment of porcine anterior segments with GAGases. ( A ) Porcine anterior segments were treated in perfusion culture with chondroitinase ABC (0.25 U/mL), heparitinase (0.01 U/mL), or hyaluronidase (10 U/mL), individually or in combination. Control eyes

    Journal: Investigative ophthalmology & visual science

    Article Title: Effects of Modifiers of Glycosaminoglycan Biosynthesis on Outflow Facility in Perfusion Culture

    doi: 10.1167/iovs.07-0903

    Figure Lengend Snippet: Treatment of porcine anterior segments with GAGases. ( A ) Porcine anterior segments were treated in perfusion culture with chondroitinase ABC (0.25 U/mL), heparitinase (0.01 U/mL), or hyaluronidase (10 U/mL), individually or in combination. Control eyes

    Article Snippet: Draq5 was obtained from Axxora (San Diego, CA); normal goat serum, mounting medium (Fluoromount G), and chamber slides (Labtek II) from Fisher Scientific (Pittsburgh, PA); chondroitinase ABC protease-free (from Proteus vulgaris ) and heparitinase (from Flavobacterium heparinum ) from Seikagaku Corp. (Tokyo, Japan); hyaluronidase (from Streptomyces hyalurolyticus ) from ICN Biomedicals (Aurora, OH); Dulbecco's modified Eagle's medium (DMEM) and penicillin-streptomycin-amphotericin B from Invitrogen (Carlsbad, CA); fetal bovine serum (FBS) from HyClone (Logan, UT); protein markers (Precision Plus) from Bio-Rad (Hercules, CA); and anti-rabbit secondary antibody (IRDye680) from Licor Biosciences (Lincoln, NE).

    Techniques:

    LMWH rescue from cell death requires HBEGF. HTR-8/SVneo cells were cultured without treatment at ambient (20%) O 2 , or exposed to H/R with no treatment (Control), supplementation with 10 IU/ml LMWH, or combinations of LMWH and either 10 μg/ml CRM197, pan-ERBB inhibitor, heparitinase I, ERBB1 blocking antibody or ERBB4 blocking antibody. HTR-8/SVneo cells were fluorescently double-labeled after treatment to visualize cell death by TUNEL (green fluorescence) and nuclei with DAPI (blue fluorescence). Representative fluorescent images are shown of TUNEL ( A – D ) or merged TUNEL/DAPI ( E – H ) in cells cultured at ambient O 2  (A, E), with H/R (B, F), with H/R + LMWH (C, G) and with H/R + LMWH + CRM197 (D, H). Bars = 100 μM. TUNEL Index is shown graphically for the indicated culture conditions and treatments, which did (solid bars) or did not (open bars) include LMWH ( I ). Error bars denote SEM. Bars labeled with dissimilar letters indicate differences at  P

    Journal: Human Reproduction (Oxford, England)

    Article Title: Enhancement of trophoblast differentiation and survival by low molecular weight heparin requires heparin-binding EGF-like growth factor

    doi: 10.1093/humrep/dex069

    Figure Lengend Snippet: LMWH rescue from cell death requires HBEGF. HTR-8/SVneo cells were cultured without treatment at ambient (20%) O 2 , or exposed to H/R with no treatment (Control), supplementation with 10 IU/ml LMWH, or combinations of LMWH and either 10 μg/ml CRM197, pan-ERBB inhibitor, heparitinase I, ERBB1 blocking antibody or ERBB4 blocking antibody. HTR-8/SVneo cells were fluorescently double-labeled after treatment to visualize cell death by TUNEL (green fluorescence) and nuclei with DAPI (blue fluorescence). Representative fluorescent images are shown of TUNEL ( A – D ) or merged TUNEL/DAPI ( E – H ) in cells cultured at ambient O 2 (A, E), with H/R (B, F), with H/R + LMWH (C, G) and with H/R + LMWH + CRM197 (D, H). Bars = 100 μM. TUNEL Index is shown graphically for the indicated culture conditions and treatments, which did (solid bars) or did not (open bars) include LMWH ( I ). Error bars denote SEM. Bars labeled with dissimilar letters indicate differences at P

    Article Snippet: Cells and villous explants were also treated with vehicle, 10 μg/ml CRM197 (a highly specific HBEGF antagonist; EMD Biosciences, San Diego, CA), 10 μg/ml EGFR/ERBB2/ERBB4 (pan-ERBB inhibitor) tyrosine kinase antagonist (Millipore, catalog no. 324 840), 0.1 U/ml heparitinase I (from Flavobacterium heparinum , EC 4.2.2.8; 100 704-1, Seikagaku America, East Falmouth, MA), and 10 μg/ml mouse anti-ERBB1 (Ab-2) or ERBB4 (Ab-3) blocking antibodies (Lab Vision, Fremont, CA).

    Techniques: Cell Culture, Blocking Assay, Labeling, TUNEL Assay, Fluorescence

    Localization of DS and HS in uninjured carotid arteries of wild-type mice. Frozen sections were treated with chondroitin B-lyase (A), Flavobacterium heparitinase (C), or buffer alone (B,D) and then incubated with monoclonal antibodies ΔDi-4S (A,B) or ΔHS (C,D). Bound monoclonal antibodies were detected with a peroxidase-conjugated secondary antibody. Arrow indicates internal elastic lamina; arrowhead, external elastic lamina. DS was present primarily in the adventitia (A) and HS in the intima/media (C).

    Journal: Blood

    Article Title: Vascular dermatan sulfate regulates the antithrombotic activity of heparin cofactor II

    doi: 10.1182/blood-2007-12-127928

    Figure Lengend Snippet: Localization of DS and HS in uninjured carotid arteries of wild-type mice. Frozen sections were treated with chondroitin B-lyase (A), Flavobacterium heparitinase (C), or buffer alone (B,D) and then incubated with monoclonal antibodies ΔDi-4S (A,B) or ΔHS (C,D). Bound monoclonal antibodies were detected with a peroxidase-conjugated secondary antibody. Arrow indicates internal elastic lamina; arrowhead, external elastic lamina. DS was present primarily in the adventitia (A) and HS in the intima/media (C).

    Article Snippet: Chondroitin B-lyase, chondroitin ABC-lyase, Flavobacterium heparitinase I, and mouse monoclonal ΔDi-4S and ΔHS antibodies were purchased from Seikagaku (Tokyo, Japan).

    Techniques: Mouse Assay, Incubation