n hydroxysuccinimide nhs  (Millipore)


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    Millipore n hydroxysuccinimide nhs
    N Hydroxysuccinimide Nhs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore biotin x nhs
    Fig. 5. The effect of thermal stress on ErbB proteins shares biochemical characteristics with the effects of CI-1033 and GA. ( A ) The indicated derivatives of 32D cells were treated for 1 h at 37°C with EGF (100 ng/ml), GA (1 µM) or CI-1033 (CI; 10 µM). Alternatively, cells were subjected to heat shock at the indicated temperature for 1 h or left untreated (lanes labeled –). Cell extracts were analyzed by IB and/or IP with the indicated antibodies. ( B ) The indicated ErbB-2 constructs were expressed in CHO cells along with HA-Ub. Forty-eight hours after transfection, cells were subjected to heat shock at 42°C for 60 min. Cell lysates were analyzed with the indicated antibodies. ( C ) T47D or T47D-5R cells were subjected to heat shock at 44°C for the indicated time intervals, followed by surface labeling for 40 min on ice with <t>Biotin-X-NHS.</t> Cell extracts were subjected to IP and IB with the indicated antibodies, or with a peroxidase-conjugated streptavidin. Note the reduction in Shc levels in the rightmost lane. ( D ) COS cells were transfected with expression vectors encoding HA-Ub and one of the four ErbB proteins. Forty-eight hours later, cells were subjected to heat stress for 60 min at 42°C as indicated (+) or left untreated (–). Following lysis, the respective ErbB proteins were analyzed with the indicated antibodies.
    Biotin X Nhs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Rate analysis of platelet removal from blood. (A) Splenic sequestration of platelets. Crushed spleens from 9/13-week-old WT and RGS18-/- mice (n = 6 per group) were stained for CD42b, and platelets were identified by flow cytometry on the basis of size and CD42b positivity. Percentages of total splenic cell population are shown. (B) Clearance of platelets. Blood cells from 7-week-old mice were <t>biotinylated</t> in vivo by infusion of <t>NHS-biotin,</t> and then blood samples were collected at the time indicated. Biotinylated platelets from WT (black line) and RGS18-/- (red line) mice (n = 9 per group) were identified by flow cytometry on whole blood using PE-Streptavidin and CD41 staining. The stability of the biotinylation was assessed by examining the in vivo biotinylated CD41 - blood cells in WT (green line) and RGS18-/- (yellow line) mice (n = 9 per group). (C) Spontaneous platelet aggregation. Whole blood cells from 9/11-week-old WT and RGS18-/- mice (n = 12 per group) were stained for CD42b, and platelets were identified by flow cytometry on the basis of size and CD42b positivity. Platelet aggregates were monitored by extending the platelet gate so as to include the platelet “smearings” having elevated FSC and FITC fluorescence. Percentages of total cell population are shown. *P
    Nhs Biotin, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore edc nhs
    a) Decrease of the resonance frequency of the quartz due to the adsorption of thiol monolayer and <t>NHS/EDC</t> cross-linker. b) Decrease of the resonance frequency of the quartz due to the adsorption of 0.1% BSA molecules.
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    Fig. 5. The effect of thermal stress on ErbB proteins shares biochemical characteristics with the effects of CI-1033 and GA. ( A ) The indicated derivatives of 32D cells were treated for 1 h at 37°C with EGF (100 ng/ml), GA (1 µM) or CI-1033 (CI; 10 µM). Alternatively, cells were subjected to heat shock at the indicated temperature for 1 h or left untreated (lanes labeled –). Cell extracts were analyzed by IB and/or IP with the indicated antibodies. ( B ) The indicated ErbB-2 constructs were expressed in CHO cells along with HA-Ub. Forty-eight hours after transfection, cells were subjected to heat shock at 42°C for 60 min. Cell lysates were analyzed with the indicated antibodies. ( C ) T47D or T47D-5R cells were subjected to heat shock at 44°C for the indicated time intervals, followed by surface labeling for 40 min on ice with Biotin-X-NHS. Cell extracts were subjected to IP and IB with the indicated antibodies, or with a peroxidase-conjugated streptavidin. Note the reduction in Shc levels in the rightmost lane. ( D ) COS cells were transfected with expression vectors encoding HA-Ub and one of the four ErbB proteins. Forty-eight hours later, cells were subjected to heat stress for 60 min at 42°C as indicated (+) or left untreated (–). Following lysis, the respective ErbB proteins were analyzed with the indicated antibodies.

    Journal: The EMBO Journal

    Article Title: Drug-induced ubiquitylation and degradation of ErbB receptor tyrosine kinases: implications for cancer therapy

    doi: 10.1093/emboj/21.10.2407

    Figure Lengend Snippet: Fig. 5. The effect of thermal stress on ErbB proteins shares biochemical characteristics with the effects of CI-1033 and GA. ( A ) The indicated derivatives of 32D cells were treated for 1 h at 37°C with EGF (100 ng/ml), GA (1 µM) or CI-1033 (CI; 10 µM). Alternatively, cells were subjected to heat shock at the indicated temperature for 1 h or left untreated (lanes labeled –). Cell extracts were analyzed by IB and/or IP with the indicated antibodies. ( B ) The indicated ErbB-2 constructs were expressed in CHO cells along with HA-Ub. Forty-eight hours after transfection, cells were subjected to heat shock at 42°C for 60 min. Cell lysates were analyzed with the indicated antibodies. ( C ) T47D or T47D-5R cells were subjected to heat shock at 44°C for the indicated time intervals, followed by surface labeling for 40 min on ice with Biotin-X-NHS. Cell extracts were subjected to IP and IB with the indicated antibodies, or with a peroxidase-conjugated streptavidin. Note the reduction in Shc levels in the rightmost lane. ( D ) COS cells were transfected with expression vectors encoding HA-Ub and one of the four ErbB proteins. Forty-eight hours later, cells were subjected to heat stress for 60 min at 42°C as indicated (+) or left untreated (–). Following lysis, the respective ErbB proteins were analyzed with the indicated antibodies.

    Article Snippet: For biotinylation experiments, cells were incubated on ice for 40 min with Biotin-X-NHS (0.5 mg/ml in PBS; Calbiochem).

    Techniques: Labeling, Construct, Transfection, Expressing, Lysis

    Fig. 3. CI-1033 and GA enhance destruction of both mature and immature ErbB-2 molecules. ( A ) Monolayers of T47D cells were incubated at 37°C for the indicated time intervals in the presence of either CI-1033 (10 µM; triangles) or Herceptin (10 µg/ml; circles). Control monolayers were incubated in the absence of agents (squares). Levels of cell surface ErbB-2 were determined by incubation with a radiolabeled antibody. Each point represents the average ± SD of duplicates. ( B ) T47D or T47D-5R cells were treated for 3 h with increasing concentrations of CI-1033 (2 or 20 µM) or with GA (1 µM). Cell extracts were subjected to IP and IB with the indicated antibodies. Note the faster mobility of the nascent ErbB-2 of T47D-5R cells. ( C ) Cells were treated with CI-1033 (5 µM) for the indicated time intervals, followed by surface labeling for 40 min on ice with Biotin-X-NHS. Streptavidin–HRP denotes incubation of the membrane with streptavidin conjugated to peroxidase. ( D ) T47D or T47D-5R cells were treated with GA (1 µM) for the indicated time intervals, followed by surface labeling for 40 min on ice with Biotin-X-NHS. Cell extracts were analyzed with the indicated antibodies.

    Journal: The EMBO Journal

    Article Title: Drug-induced ubiquitylation and degradation of ErbB receptor tyrosine kinases: implications for cancer therapy

    doi: 10.1093/emboj/21.10.2407

    Figure Lengend Snippet: Fig. 3. CI-1033 and GA enhance destruction of both mature and immature ErbB-2 molecules. ( A ) Monolayers of T47D cells were incubated at 37°C for the indicated time intervals in the presence of either CI-1033 (10 µM; triangles) or Herceptin (10 µg/ml; circles). Control monolayers were incubated in the absence of agents (squares). Levels of cell surface ErbB-2 were determined by incubation with a radiolabeled antibody. Each point represents the average ± SD of duplicates. ( B ) T47D or T47D-5R cells were treated for 3 h with increasing concentrations of CI-1033 (2 or 20 µM) or with GA (1 µM). Cell extracts were subjected to IP and IB with the indicated antibodies. Note the faster mobility of the nascent ErbB-2 of T47D-5R cells. ( C ) Cells were treated with CI-1033 (5 µM) for the indicated time intervals, followed by surface labeling for 40 min on ice with Biotin-X-NHS. Streptavidin–HRP denotes incubation of the membrane with streptavidin conjugated to peroxidase. ( D ) T47D or T47D-5R cells were treated with GA (1 µM) for the indicated time intervals, followed by surface labeling for 40 min on ice with Biotin-X-NHS. Cell extracts were analyzed with the indicated antibodies.

    Article Snippet: For biotinylation experiments, cells were incubated on ice for 40 min with Biotin-X-NHS (0.5 mg/ml in PBS; Calbiochem).

    Techniques: Incubation, Labeling

    Rate analysis of platelet removal from blood. (A) Splenic sequestration of platelets. Crushed spleens from 9/13-week-old WT and RGS18-/- mice (n = 6 per group) were stained for CD42b, and platelets were identified by flow cytometry on the basis of size and CD42b positivity. Percentages of total splenic cell population are shown. (B) Clearance of platelets. Blood cells from 7-week-old mice were biotinylated in vivo by infusion of NHS-biotin, and then blood samples were collected at the time indicated. Biotinylated platelets from WT (black line) and RGS18-/- (red line) mice (n = 9 per group) were identified by flow cytometry on whole blood using PE-Streptavidin and CD41 staining. The stability of the biotinylation was assessed by examining the in vivo biotinylated CD41 - blood cells in WT (green line) and RGS18-/- (yellow line) mice (n = 9 per group). (C) Spontaneous platelet aggregation. Whole blood cells from 9/11-week-old WT and RGS18-/- mice (n = 12 per group) were stained for CD42b, and platelets were identified by flow cytometry on the basis of size and CD42b positivity. Platelet aggregates were monitored by extending the platelet gate so as to include the platelet “smearings” having elevated FSC and FITC fluorescence. Percentages of total cell population are shown. *P

    Journal: PLoS ONE

    Article Title: Regulator of G-Protein Signaling 18 Controls Both Platelet Generation and Function

    doi: 10.1371/journal.pone.0113215

    Figure Lengend Snippet: Rate analysis of platelet removal from blood. (A) Splenic sequestration of platelets. Crushed spleens from 9/13-week-old WT and RGS18-/- mice (n = 6 per group) were stained for CD42b, and platelets were identified by flow cytometry on the basis of size and CD42b positivity. Percentages of total splenic cell population are shown. (B) Clearance of platelets. Blood cells from 7-week-old mice were biotinylated in vivo by infusion of NHS-biotin, and then blood samples were collected at the time indicated. Biotinylated platelets from WT (black line) and RGS18-/- (red line) mice (n = 9 per group) were identified by flow cytometry on whole blood using PE-Streptavidin and CD41 staining. The stability of the biotinylation was assessed by examining the in vivo biotinylated CD41 - blood cells in WT (green line) and RGS18-/- (yellow line) mice (n = 9 per group). (C) Spontaneous platelet aggregation. Whole blood cells from 9/11-week-old WT and RGS18-/- mice (n = 12 per group) were stained for CD42b, and platelets were identified by flow cytometry on the basis of size and CD42b positivity. Platelet aggregates were monitored by extending the platelet gate so as to include the platelet “smearings” having elevated FSC and FITC fluorescence. Percentages of total cell population are shown. *P

    Article Snippet: Platelet Clearance Mice platelets were biotinylated in vivo by infusion with NHS-biotin (Calbiochem, La Jolla, CA, USA).

    Techniques: Mouse Assay, Staining, Flow Cytometry, Cytometry, In Vivo, Fluorescence

    Surface biotinylation of MDCK-GP and MBGV-infected MDCKII cells. (A) MDCK-GP and MDCKII (Mock) cells were grown for 48 h on membrane filters (24-mm diameter). Then, the cells were incubated with NHS-biotin from either the apical (lanes a) or the basolateral (lanes b) side. After lysis of the cells, GP was immunoprecipitated using a rabbit anti-GP antibody, and immunocomplexes were subjected to SDS-PAGE and Western blot. Biotinylated proteins were detected with peroxidase-coupled streptavidin followed by chemiluminescence analysis. (B) MDCKII cells in suspension were infected with MBGV at an MOI of 1 PFU per cell or left untreated (Mock) and cultivated on membrane filters for 48 h. Polarized cells were subjected to surface biotinylation as described above.

    Journal: Journal of Virology

    Article Title: Sorting of Marburg Virus Surface Protein and Virus Release Take Place at Opposite Surfaces of Infected Polarized Epithelial Cells

    doi: 10.1128/JVI.75.3.1274-1283.2001

    Figure Lengend Snippet: Surface biotinylation of MDCK-GP and MBGV-infected MDCKII cells. (A) MDCK-GP and MDCKII (Mock) cells were grown for 48 h on membrane filters (24-mm diameter). Then, the cells were incubated with NHS-biotin from either the apical (lanes a) or the basolateral (lanes b) side. After lysis of the cells, GP was immunoprecipitated using a rabbit anti-GP antibody, and immunocomplexes were subjected to SDS-PAGE and Western blot. Biotinylated proteins were detected with peroxidase-coupled streptavidin followed by chemiluminescence analysis. (B) MDCKII cells in suspension were infected with MBGV at an MOI of 1 PFU per cell or left untreated (Mock) and cultivated on membrane filters for 48 h. Polarized cells were subjected to surface biotinylation as described above.

    Article Snippet: Either the apical or the basolateral membrane of filter-grown polarized MDCK-GP cells was labeled with NHS-biotin under conditions which allow the specific detection of surface proteins.

    Techniques: Infection, Incubation, Lysis, Immunoprecipitation, SDS Page, Western Blot

    a) Decrease of the resonance frequency of the quartz due to the adsorption of thiol monolayer and NHS/EDC cross-linker. b) Decrease of the resonance frequency of the quartz due to the adsorption of 0.1% BSA molecules.

    Journal: Sensors (Basel, Switzerland)

    Article Title: Electrical Characterization of a Thiol SAM on Gold as a First Step for the Fabrication of Immunosensors based on a Quartz Crystal Microbalance

    doi:

    Figure Lengend Snippet: a) Decrease of the resonance frequency of the quartz due to the adsorption of thiol monolayer and NHS/EDC cross-linker. b) Decrease of the resonance frequency of the quartz due to the adsorption of 0.1% BSA molecules.

    Article Snippet: Activation with EDC/NHS 0.5 M NHS and 0.2 M EDC solution have been prepared in millipore water.

    Techniques: Adsorption