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5c2  (Novus Biologicals)


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    Structured Review

    Novus Biologicals 5c2
    5c2, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/5c2/alpha-Synuclein+Antibody+(5C2)+-+BSA+Free/pm40301692-411-52-53
    Average 93 stars, based on 7 article reviews
    5c2 - by Bioz Stars, 2026-09
    93/100 stars

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    Incubation:

    Article Title: Flow cytometric isolation of drug-like conformational antibodies specific for amyloid fibrils.
    Article Snippet: .. 13 Tau dot blots, as reported in Fig. S6A, were next incubated with antibodies at 10 nM (1% nonfat dry milk in 14 TBST) overnight at 4 °C. α-synuclein dot blots, as reported in Fig. S14, were incubated with 50 nM antibody 15 (aS2.1 WT, H2.7, H2.4, H2.3) or a 1000x dilution of 5C2 (Novus Biologicals, NBP1-04321), LB509 (Abcam, 16 ab27766) and MJFR1 (Abcam, ab2099538). .. Next, the blots were washed with TBST and incubated with a 5000x 17 diluted solution of HRP-conjugated goat anti-human IgG or a 1000x diluted solution of HRP-conjugated goat 18 anti-mouse or goat anti-rabbit at room temperature for 1 h. Afterward, the blots were washed with TBST and 19 developed using Ecobright Pico HRP Substrate (Innovative Solutions) and visualized with the Genesys G:Box 20 imaging system (Syngene).

    Article Title: Facile generation of drug-like conformational antibodies specific for amyloid fibrils.
    Article Snippet: Antibodies that recognize insoluble antigens, such as amyloid fibrils associated with neurodegenerative disorders, are important for research, diagnostic and therapeutic applications.. However, these types of antibodies are difficult to generate, typically require animal immunization and also commonly require humanization in the case of therapeutic applications.. Here we report a methodology for generating high-quality, fully human, conformation-specific antibodies against amyloid fibrils using a published human nonimmune library, yeast-surface display and quantitative fluorescence-activated cell sorting.

    Article Title: Flow cytometric isolation of drug-like conformational antibodies specific for amyloid fibrils
    Article Snippet: .. Tau dot blots, as reported in Fig. S6A , were next incubated with antibodies at 10 nM (1% nonfat dry milk in TBST) overnight at 4 °C. α-synuclein dot blots, as reported in Fig. S14 , were incubated with 50 nM antibody (aS2.1 WT, H2.7, H2.4, H2.3) or a 1000x dilution of 5C2 (Novus Biologicals, NBP1-04321), LB509 (Abcam, ab27766) and MJFR1 (Abcam, ab2099538). .. Next, the blots were washed with TBST and incubated with a 5000x diluted solution of HRP-conjugated goat anti-human IgG or a 1000x diluted solution of HRP-conjugated goat anti-mouse or goat anti-rabbit at room temperature for 1 h. Afterward, the blots were washed with TBST and developed using Ecobright Pico HRP Substrate (Innovative Solutions) and visualized with the Genesys G:Box imaging system (Syngene).

    Binding Assay:

    Article Title: Nature-inspired design and evolution of anti-amyloid antibodies
    Article Snippet: .. Antibody binding was performed by incubating the blocked membranes with 10 ml of 100 n m scFv, 10 n m scFv–Fc fusion, 1:1000 dilution of NAB228 (2 mg/ml stock concentration; A8354, Sigma), 1E1/A6 (stock concentration unknown; 05-804, EMD Millipore), or 1:10,000 of 5C2 (1 mg/ml stock concentration; NBP1-04321, Novus Biologicals). ..

    Article Title: Nature-inspired design and evolution of anti-amyloid antibodies
    Article Snippet: .. Antibody binding was performed by incubating the blocked membranes with 10 mL of 100 nM scFv, 10 nM scFv-Fc fusion, 1:1000 dilution of NAB228 (2 mg/mL stock concentration; A8354, Sigma-Aldrich), 1E1/A6 (stock concentration unknown; 05-804, EMD Millipore) or 1:10000 of 5C2 (1 mg/mL stock concentration; NBP1-04321, Novus Biologicals). ..

    Concentration Assay:

    Article Title: Nature-inspired design and evolution of anti-amyloid antibodies
    Article Snippet: .. Antibody binding was performed by incubating the blocked membranes with 10 ml of 100 n m scFv, 10 n m scFv–Fc fusion, 1:1000 dilution of NAB228 (2 mg/ml stock concentration; A8354, Sigma), 1E1/A6 (stock concentration unknown; 05-804, EMD Millipore), or 1:10,000 of 5C2 (1 mg/ml stock concentration; NBP1-04321, Novus Biologicals). ..

    Article Title: Nature-inspired design and evolution of anti-amyloid antibodies
    Article Snippet: .. Antibody binding was performed by incubating the blocked membranes with 10 mL of 100 nM scFv, 10 nM scFv-Fc fusion, 1:1000 dilution of NAB228 (2 mg/mL stock concentration; A8354, Sigma-Aldrich), 1E1/A6 (stock concentration unknown; 05-804, EMD Millipore) or 1:10000 of 5C2 (1 mg/mL stock concentration; NBP1-04321, Novus Biologicals). ..

    Western Blot:

    Article Title: Anti-amyloid Compounds Inhibit α-Synuclein Aggregation Induced by Protein Misfolding Cyclic Amplification (PMCA)
    Article Snippet: .. The anti-α-synuclein antibodies Syn1 (BD Biosciences), 5C2 (Novus Biologicals), Per7 ( ) and Per4 ( ) were used for immunoblotting. ..

    Generated:

    Article Title: Discrimination of MSA-P and MSA-C by RT-QuIC analysis of olfactory mucosa: the first assessment of assay reproducibility between two specialized laboratories
    Article Snippet: .. Eight μL of αSyn_RT-QuIC products generated by MSA-P and MSA-C samples were treated with PK [2.5 mg/mL] for 1 h at 37 °C under shaking (500 rpm) and immunoblotted with antibodies directed against three different epitopes of α-synuclein: (i) clone 5C2 (epitopes 61–95, Novus Biologicals), (ii) AB5038 (epitopes 111–131, Chemicon international), and (iii) clone 42 (epitopes 15–123, BD Bioscience). ..



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    Fig. 5. <t>ATM</t> interacts with CRMP5 and Spastin (SPAST) in vitro. (A) Immunoprecipitation of ATM <t>by</t> <t>anti-ATM</t> (5C2) antibody in unstressed and Taxol-treated cells, with detection of CRMP5 and SPAST as interacting proteins. (B) Immunoprecipitation of CRMP5 protein in unstressed and Taxol-treated cells, with detection of ATM and SPAST as interacting proteins. Abbreviations: SPAST = Spastin.
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    Image Search Results


    Fig. 5. ATM interacts with CRMP5 and Spastin (SPAST) in vitro. (A) Immunoprecipitation of ATM by anti-ATM (5C2) antibody in unstressed and Taxol-treated cells, with detection of CRMP5 and SPAST as interacting proteins. (B) Immunoprecipitation of CRMP5 protein in unstressed and Taxol-treated cells, with detection of ATM and SPAST as interacting proteins. Abbreviations: SPAST = Spastin.

    Journal: Neurobiology of disease

    Article Title: The ataxia-telangiectasia disease protein ATM controls vesicular protein secretion via CHGA and microtubule dynamics via CRMP5.

    doi: 10.1016/j.nbd.2024.106756

    Figure Lengend Snippet: Fig. 5. ATM interacts with CRMP5 and Spastin (SPAST) in vitro. (A) Immunoprecipitation of ATM by anti-ATM (5C2) antibody in unstressed and Taxol-treated cells, with detection of CRMP5 and SPAST as interacting proteins. (B) Immunoprecipitation of CRMP5 protein in unstressed and Taxol-treated cells, with detection of ATM and SPAST as interacting proteins. Abbreviations: SPAST = Spastin.

    Article Snippet: For co-immunoprecipitations, 800–1000 μg protein from cytoplasmic subcellular fractions of SH-SY5Y cells were incubated with either 10 μg anti-ATM antibody (Novus Biologicals, Littleton, Colorado, USA, NB100–220) or 10 μg of mouse IgG1 isotype control antibody (Cell Signaling Technology, #5415) in binding buffer consisting of CEB fractionation buffer supplemented with HALT phosphatase inhibitors (Thermo Fisher Scientific) and cOmplete protease inhibitors (Roche) overnight at 4 ◦C with head to tail rotation.

    Techniques: In Vitro, Immunoprecipitation

    Fig. 8. Proposed pathway of novel cytoplasmic ATM-mediated signaling events. CHGA protein levels and secretion is reduced compared to normal cells with neural origin (right panel) and CRMP5 protein levels as well as phosphorylation at Ser538 are diminished upon ATM loss (left panel). Given the crucial role of CHGA in dense-core granule biogenesis (Koshimizu et al., 2010), we propose that reduced CHGA in granules in the trans-Golgi network (TGN) diminishes budding of large dense core vesicles (LDCV) from TGN membrane resulting in reduced transport of LDCVs along the axon and reduced secretion of LDCVs to the extracellular space. This likely results in reduced neuropeptide and neurotrophic signaling. Concomitant CRMP5 protein loss causes increased polymerization of microtubules, probably via reduced binding and sequestration of tubulin heterodimers, shifting the microtubule equilibrium from dynamic towards stabilized, thereby causing neurite retraction. Abbreviations: CHGA = Chromogranin A; CRMP5 = Collapsin Response Mediator Protein 5; ER = endoplasmic reticulum; INA = Internexin-alpha; LDCV = large dense-core vesicle; NFL = Neurofilament light chain; TANC2 = Tetratricopeptide repeat, ankyrin repeat and coiled-coil containing 2 protein.

    Journal: Neurobiology of disease

    Article Title: The ataxia-telangiectasia disease protein ATM controls vesicular protein secretion via CHGA and microtubule dynamics via CRMP5.

    doi: 10.1016/j.nbd.2024.106756

    Figure Lengend Snippet: Fig. 8. Proposed pathway of novel cytoplasmic ATM-mediated signaling events. CHGA protein levels and secretion is reduced compared to normal cells with neural origin (right panel) and CRMP5 protein levels as well as phosphorylation at Ser538 are diminished upon ATM loss (left panel). Given the crucial role of CHGA in dense-core granule biogenesis (Koshimizu et al., 2010), we propose that reduced CHGA in granules in the trans-Golgi network (TGN) diminishes budding of large dense core vesicles (LDCV) from TGN membrane resulting in reduced transport of LDCVs along the axon and reduced secretion of LDCVs to the extracellular space. This likely results in reduced neuropeptide and neurotrophic signaling. Concomitant CRMP5 protein loss causes increased polymerization of microtubules, probably via reduced binding and sequestration of tubulin heterodimers, shifting the microtubule equilibrium from dynamic towards stabilized, thereby causing neurite retraction. Abbreviations: CHGA = Chromogranin A; CRMP5 = Collapsin Response Mediator Protein 5; ER = endoplasmic reticulum; INA = Internexin-alpha; LDCV = large dense-core vesicle; NFL = Neurofilament light chain; TANC2 = Tetratricopeptide repeat, ankyrin repeat and coiled-coil containing 2 protein.

    Article Snippet: For co-immunoprecipitations, 800–1000 μg protein from cytoplasmic subcellular fractions of SH-SY5Y cells were incubated with either 10 μg anti-ATM antibody (Novus Biologicals, Littleton, Colorado, USA, NB100–220) or 10 μg of mouse IgG1 isotype control antibody (Cell Signaling Technology, #5415) in binding buffer consisting of CEB fractionation buffer supplemented with HALT phosphatase inhibitors (Thermo Fisher Scientific) and cOmplete protease inhibitors (Roche) overnight at 4 ◦C with head to tail rotation.

    Techniques: Phospho-proteomics, Membrane, Binding Assay