endoglycosidase treatment  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Name:
    Endoglycosidase Reaction Buffer Pack
    Description:
    Endoglycosidase Reaction Buffer Pack 4 0 ml
    Catalog Number:
    b0701s
    Price:
    23
    Size:
    4 0 ml
    Category:
    Buffers
    Buy from Supplier


    Structured Review

    New England Biolabs endoglycosidase treatment
    Endoglycosidase Reaction Buffer Pack
    Endoglycosidase Reaction Buffer Pack 4 0 ml
    https://www.bioz.com/result/endoglycosidase treatment/product/New England Biolabs
    Average 90 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    endoglycosidase treatment - by Bioz Stars, 2020-08
    90/100 stars

    Images

    Related Articles

    Incubation:

    Article Title: The C-terminal Cytosolic Region of Rim21 Senses Alterations in Plasma Membrane Lipid Composition
    Article Snippet: .. The prepared total cell lysates were diluted with four volumes of endoglycosidase H buffer (62.5 m m sodium citrate, pH 5.5, and 1.25 m m PMSF) and incubated with 20 units/μl endoglycosidase H (Endo Hf ; New England Biolabs, Beverly, MA) at 37 °C for 1 h with occasional mixing by tapping. ..

    Article Title: N-glycosylation of ICAM-2 is required for ICAM-2-mediated complete suppression of metastatic potential of SK-N-AS neuroblastoma cells
    Article Snippet: .. After cooling, protein solutions were centrifuged and incubated for an additional hour at 37°C with 2 μl of either Endo H enzyme (plus G5 Reaction buffer, pH 5.5; New England Biolabs) or PNGase F enzyme (plus G7 reaction buffer, pH 7.5 plus 10% NP-40 ; New England Biolabs). .. Samples were then subjected to SDS-PAGE and immunoblot analyses.

    Article Title: Ambroxol improves lysosomal biochemistry in glucocerebrosidase mutation-linked Parkinson disease cells
    Article Snippet: .. The lysate was then incubated for 12 h with 1 000 units of endoglycosidase-H and reaction buffer (New England Biolabs 0.5 M sodium citrate, pH 5.5). ..

    Article Title: Post-Translational Processing of Synaptophysin in the Rat Retina Is Disrupted by Diabetes
    Article Snippet: .. Endo H Treatment of Retinal Lysates 6 µl of 1X denaturing buffer (New England Biolabs, Ipswich, MA, USA) was added to 30 µg of retinal lysate and boiled for 7 min. 15 µg was taken from each sample and incubated with endoglycosidase H (endo H) and 1X ‘G5’ reaction buffer (New England Biolabs, Ipswich, MA, USA) for 1 hr at 37°C. .. The reaction was stopped by the addition of sample buffer (1X NuPAGE® LDS, Invitrogen®, California, USA) and heated at 70°C for 15 min.

    Article Title: Pharmacological Chaperones and Coenzyme Q10 Treatment Improves Mutant β-Glucocerebrosidase Activity and Mitochondrial Function in Neuronopathic Forms of Gaucher Disease
    Article Snippet: .. The lysate was subjected to an overnight incubation with endoglycosidase-H and reaction buffer (New England Biolabs 0.5 M sodium citrate, pH 5.5) according to the manufacturer’s instructions. .. The lysates were then analyzed by Western blot with the anti GCase antibody.

    SDS Page:

    Article Title: Re-evaluation of neuronal P2X7 expression using novel mouse models and a P2X7-specific nanobody
    Article Snippet: .. 10ml of extract were separated by SDS-PAGE with with or without endoglycosidase treatment (30 min at 37 °C in the presence of reducing loading buffer (1x) and 5 IUB miliunits EndoH or 10 IUB miliunits PNGaseF (New England Biolabs)). .. For ethidium uptake measurements, cells were seeded after 27 h in 96-well plates (5x104 cells/well) and incubated in the presence of 20 mM ethidium bromide in PBS for 15 min. Dye influx was evaluated with a fluorescence plate reader (Fluostar Galaxy, BMG) upon addition of the indicated ATP concentrations, as described ( ).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    New England Biolabs endoglycosidase treatment
    Expression and functionality of the P2X7-EGFP constructs in HEK cells and expression of the transgene in mice. ( A ) The EGFP sequence was fused via a Strep-tag-His-tag linker to the very C-terminus of the mouse P2X7 sequence. (B/C) Protein extracts from transiently transfected (Lipofectamine 2000, Invitrogen) HEK cells (DSMZ (ACC 305), regularly tested for mycoplasma contamination) were separated by SDS-PAGE with <t>endoglycosidase</t> treatment as indicated. Gels were analyzed by western blotting with a P2X7-specific antibody (Alamone, extracellular) ( B ) or by direct EGFP-fluorescence scanning ( C ). ( D ) Normalized dose–response curves for ATP-induced ethidium uptake. HEK293 cells were cultured and transfected (2 μg DNA/well of a six-well-plate, Lipofectamin, Thermo Fisher Scientific). After 27 hr, cells were seeded in 96-well plates (5 × 10 4 cells/well) and incubated in the presence of 20 μM ethidium bromide in PBS for 15 min. Dye influx was evaluated with a fluorescence plate reader (Fluostar Galaxy, BMG) upon addition of the indicated ATP concentrations, as described ( Bruzzone et al., 2010 ). Lines represent nonlinear curve fits of the Hill equation to the data and were normalized to the calculated maximal responses. EC 50 values are 582 (CI 498–681), 840 (CI 644–1098) and 582 (CI 457–740) for wt (L variant) and EGFP-tagged L and P variants, respectively. Error bars represent SEM (n = 4–7). CI = 95% confidence interval. ( E ) Patch-clamp recordings from HEK cells transfected as above with wt and EGFP-tagged P2X7 (L variants). Recordings were performed as described ( Nicke et al., 2009 ) in normal or low divalent cation (DIC) containing extracellular solution to account for possible unspecific effects ( Nörenberg et al., 2016 ). Representative current traces from n > 3 cells are shown. Due to problems with the perfusion, one trace is incomplete (1 mM ATP for P2X7-EGFP in low DIC). ( F ) Southern blot controls for correct modification and integrity of the P2X7 BAC clone during subsequent homologous recombination steps. ( G ) Reproducibility and stability of endogenous and transgenic P2X7 protein expression. Protein extracts from four line 17 mice were separated by SDS-PAGE and quantified by western blotting and infrared imaging using antibodies against P2X7 (Synaptic Systems) and vinculin and fluorescent secondary antibodies. Data are presented as mean ±SD from four individual mice. No significant difference of endogenous P2X7 expression was found in two independent experiments.
    Endoglycosidase Treatment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/endoglycosidase treatment/product/New England Biolabs
    Average 90 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    endoglycosidase treatment - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    Image Search Results


    Expression and functionality of the P2X7-EGFP constructs in HEK cells and expression of the transgene in mice. ( A ) The EGFP sequence was fused via a Strep-tag-His-tag linker to the very C-terminus of the mouse P2X7 sequence. (B/C) Protein extracts from transiently transfected (Lipofectamine 2000, Invitrogen) HEK cells (DSMZ (ACC 305), regularly tested for mycoplasma contamination) were separated by SDS-PAGE with endoglycosidase treatment as indicated. Gels were analyzed by western blotting with a P2X7-specific antibody (Alamone, extracellular) ( B ) or by direct EGFP-fluorescence scanning ( C ). ( D ) Normalized dose–response curves for ATP-induced ethidium uptake. HEK293 cells were cultured and transfected (2 μg DNA/well of a six-well-plate, Lipofectamin, Thermo Fisher Scientific). After 27 hr, cells were seeded in 96-well plates (5 × 10 4 cells/well) and incubated in the presence of 20 μM ethidium bromide in PBS for 15 min. Dye influx was evaluated with a fluorescence plate reader (Fluostar Galaxy, BMG) upon addition of the indicated ATP concentrations, as described ( Bruzzone et al., 2010 ). Lines represent nonlinear curve fits of the Hill equation to the data and were normalized to the calculated maximal responses. EC 50 values are 582 (CI 498–681), 840 (CI 644–1098) and 582 (CI 457–740) for wt (L variant) and EGFP-tagged L and P variants, respectively. Error bars represent SEM (n = 4–7). CI = 95% confidence interval. ( E ) Patch-clamp recordings from HEK cells transfected as above with wt and EGFP-tagged P2X7 (L variants). Recordings were performed as described ( Nicke et al., 2009 ) in normal or low divalent cation (DIC) containing extracellular solution to account for possible unspecific effects ( Nörenberg et al., 2016 ). Representative current traces from n > 3 cells are shown. Due to problems with the perfusion, one trace is incomplete (1 mM ATP for P2X7-EGFP in low DIC). ( F ) Southern blot controls for correct modification and integrity of the P2X7 BAC clone during subsequent homologous recombination steps. ( G ) Reproducibility and stability of endogenous and transgenic P2X7 protein expression. Protein extracts from four line 17 mice were separated by SDS-PAGE and quantified by western blotting and infrared imaging using antibodies against P2X7 (Synaptic Systems) and vinculin and fluorescent secondary antibodies. Data are presented as mean ±SD from four individual mice. No significant difference of endogenous P2X7 expression was found in two independent experiments.

    Journal: eLife

    Article Title: Re-evaluation of neuronal P2X7 expression using novel mouse models and a P2X7-specific nanobody

    doi: 10.7554/eLife.36217

    Figure Lengend Snippet: Expression and functionality of the P2X7-EGFP constructs in HEK cells and expression of the transgene in mice. ( A ) The EGFP sequence was fused via a Strep-tag-His-tag linker to the very C-terminus of the mouse P2X7 sequence. (B/C) Protein extracts from transiently transfected (Lipofectamine 2000, Invitrogen) HEK cells (DSMZ (ACC 305), regularly tested for mycoplasma contamination) were separated by SDS-PAGE with endoglycosidase treatment as indicated. Gels were analyzed by western blotting with a P2X7-specific antibody (Alamone, extracellular) ( B ) or by direct EGFP-fluorescence scanning ( C ). ( D ) Normalized dose–response curves for ATP-induced ethidium uptake. HEK293 cells were cultured and transfected (2 μg DNA/well of a six-well-plate, Lipofectamin, Thermo Fisher Scientific). After 27 hr, cells were seeded in 96-well plates (5 × 10 4 cells/well) and incubated in the presence of 20 μM ethidium bromide in PBS for 15 min. Dye influx was evaluated with a fluorescence plate reader (Fluostar Galaxy, BMG) upon addition of the indicated ATP concentrations, as described ( Bruzzone et al., 2010 ). Lines represent nonlinear curve fits of the Hill equation to the data and were normalized to the calculated maximal responses. EC 50 values are 582 (CI 498–681), 840 (CI 644–1098) and 582 (CI 457–740) for wt (L variant) and EGFP-tagged L and P variants, respectively. Error bars represent SEM (n = 4–7). CI = 95% confidence interval. ( E ) Patch-clamp recordings from HEK cells transfected as above with wt and EGFP-tagged P2X7 (L variants). Recordings were performed as described ( Nicke et al., 2009 ) in normal or low divalent cation (DIC) containing extracellular solution to account for possible unspecific effects ( Nörenberg et al., 2016 ). Representative current traces from n > 3 cells are shown. Due to problems with the perfusion, one trace is incomplete (1 mM ATP for P2X7-EGFP in low DIC). ( F ) Southern blot controls for correct modification and integrity of the P2X7 BAC clone during subsequent homologous recombination steps. ( G ) Reproducibility and stability of endogenous and transgenic P2X7 protein expression. Protein extracts from four line 17 mice were separated by SDS-PAGE and quantified by western blotting and infrared imaging using antibodies against P2X7 (Synaptic Systems) and vinculin and fluorescent secondary antibodies. Data are presented as mean ±SD from four individual mice. No significant difference of endogenous P2X7 expression was found in two independent experiments.

    Article Snippet: 10ml of extract were separated by SDS-PAGE with with or without endoglycosidase treatment (30 min at 37 °C in the presence of reducing loading buffer (1x) and 5 IUB miliunits EndoH or 10 IUB miliunits PNGaseF (New England Biolabs)).

    Techniques: Expressing, Construct, Mouse Assay, Sequencing, Strep-tag, Transfection, SDS Page, Western Blot, Fluorescence, Cell Culture, Incubation, Variant Assay, Patch Clamp, Southern Blot, Modification, BAC Assay, Homologous Recombination, Transgenic Assay, Imaging