streptavidin conjugated to alkaline phosphatase  (GE Healthcare)

 
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    GE Healthcare streptavidin conjugated to alkaline phosphatase
    Streptavidin Conjugated To Alkaline Phosphatase, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 96/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin conjugated to alkaline phosphatase/product/GE Healthcare
    Average 96 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    streptavidin conjugated to alkaline phosphatase - by Bioz Stars, 2020-09
    96/100 stars

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    Incubation:

    Article Title: Generation of Antibody-Producing Hybridomas Following One Single Immunization with a Targeted DNA Vaccine
    Article Snippet: .. Samples (50 μl/well) were incubated overnight followed by incubation with 75 μl/well of biotinylated detection antibodies [clone 1 or 2 (1 μg/ml) or class or subclass specific antibodies] (incubation time approximately 1.5 h) and alkaline phosphatase-conjugated streptavidin (1:3000, approximately 1.5 h) (GE Healthcare). .. P-nitrophenyl phosphate in diethanolamine buffer was developed for 10–60 min, and the absorbance was measured at 405 nm with a Tecan Sunrise Microplate Reader (Tecan Austria Gesellschaft, Salzburg, Austria).

    Article Title: Poly(ADP-Ribose) Polymerase 1 Binds to Kaposi's Sarcoma-Associated Herpesvirus (KSHV) Terminal Repeat Sequence and Modulates KSHV Replication in Latency
    Article Snippet: .. The wells were washed and incubated with streptavidin conjugated with alkaline phosphatase (streptavidin-AP) (Amersham Bioscience) in PBS-T for 30 min at RT, and then the bound streptavidin-AP was detected colorimetrically with p -nitrophenyl phosphate (pNpp) tablets (Bio-Rad) and a diethanolamine buffer according to the manufacturer's protocol (Bio-Rad). .. The color intensity was measured with an absorbance meter (Benchmark; Bio-Rad) at 405 nm.

    Article Title: Receptors and Lethal Effect of Bacillus thuringiensis Insecticidal Crystal Proteins to the Anticarsia gemmatalis (Lepidoptera, Noctuidae)
    Article Snippet: .. The tissue sections were then incubated with 1.5 to 6 μ g of biotinylated toxins (Cry1Aa, Cry1Ac, Cry1Ba, Cry1Da, Cry1Ea, and Cry2Aa) in the TST-buffer for 1 h. Following a washing step with the TST-buffer, the tissues were covered with 300 μ L of streptavidin-alkaline phosphatase conjugate (Amersham) diluted 1/300 in TST-buffer. .. Following a 1 h incubation period, unbound streptavidin-enzyme conjugate was removed by washing with the TST-buffer and the bound toxin was finally visualized by incubation with an alkaline phosphatase substrate solution (1.75 mg of 5-bromo-4-chloro-3-indolyl phosphate and 2.5 mg of nitroblue tetrazolium in 10 mL of buffer containing 100 mM Tris, 100 mM NaCl, and 5 mM MgCl2 ; pH 9.5) or a peroxidase substrate (0.01% 3,34-diaminobenzidine, 0.003% H2 O2 in 50 mM Tris; pH 7.6).

    Labeling:

    Article Title: TGF?1 signaling via ?V?6 integrin
    Article Snippet: .. To visualize all transferred proteins, we used the ECL protein biotinylation labeling modules (RPN 2202, Amersham) and streptavidin alkaline phosphatase (V020402, Amersham) in accordance with the manufacturer's instructions. .. Ras activation assay Only activated p21Ras is able to bind Raf1, leading to a Raf1-translocation to the cell membrane.

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    GE Healthcare alkaline phosphatase ap conjugated streptavidin
    Inhibition of mAb binding to HMW-MAA by synthetic peptides. Microtiter plates were coated with mAb TP61.5 and incubated with microsomal preparation of 518A2 melanoma cells to catch HMW-MAA. Biotinylated mAbs were preincubated with increasing concentrations of synthetic peptides, followed by incubation with HMW-MAA. ( A ) Biotinylated mAb VT80.12 was preincubated with peptide 15/3/6 (•) or an irrelevant control peptide (▴). ( B ) Biotinylated mAb VF1-TP43 was preincubated with peptide 43.12p3 (▪) as well as the control peptide (▴). Binding of biotinylated mAbs was measured using an AP-conjugated <t>streptavidin.</t> Percentage of inhibition was calculated as follows: 100−(OD (inhibited)/OD (uninhibited)×100).
    Alkaline Phosphatase Ap Conjugated Streptavidin, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 96/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alkaline phosphatase ap conjugated streptavidin/product/GE Healthcare
    Average 96 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase ap conjugated streptavidin - by Bioz Stars, 2020-09
    96/100 stars
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    Inhibition of mAb binding to HMW-MAA by synthetic peptides. Microtiter plates were coated with mAb TP61.5 and incubated with microsomal preparation of 518A2 melanoma cells to catch HMW-MAA. Biotinylated mAbs were preincubated with increasing concentrations of synthetic peptides, followed by incubation with HMW-MAA. ( A ) Biotinylated mAb VT80.12 was preincubated with peptide 15/3/6 (•) or an irrelevant control peptide (▴). ( B ) Biotinylated mAb VF1-TP43 was preincubated with peptide 43.12p3 (▪) as well as the control peptide (▴). Binding of biotinylated mAbs was measured using an AP-conjugated streptavidin. Percentage of inhibition was calculated as follows: 100−(OD (inhibited)/OD (uninhibited)×100).

    Journal: PLoS ONE

    Article Title: Specificity of Mimotope-Induced Anti-High Molecular Weight-Melanoma Associated Antigen (HMW-MAA) Antibodies Does Not Ensure Biological Activity

    doi: 10.1371/journal.pone.0019383

    Figure Lengend Snippet: Inhibition of mAb binding to HMW-MAA by synthetic peptides. Microtiter plates were coated with mAb TP61.5 and incubated with microsomal preparation of 518A2 melanoma cells to catch HMW-MAA. Biotinylated mAbs were preincubated with increasing concentrations of synthetic peptides, followed by incubation with HMW-MAA. ( A ) Biotinylated mAb VT80.12 was preincubated with peptide 15/3/6 (•) or an irrelevant control peptide (▴). ( B ) Biotinylated mAb VF1-TP43 was preincubated with peptide 43.12p3 (▪) as well as the control peptide (▴). Binding of biotinylated mAbs was measured using an AP-conjugated streptavidin. Percentage of inhibition was calculated as follows: 100−(OD (inhibited)/OD (uninhibited)×100).

    Article Snippet: Bound biotinylated mAb was detected using alkaline phosphatase (AP)-conjugated streptavidin (GE Healthcare, Little Chalfont, UK), followed by the addition of p-nitrophenylphosphate (Sigma).

    Techniques: Inhibition, Binding Assay, Incubation

    Specificity of the anti-mCherry Abs clone 1 and 2. (A) Microscopy revealed that the anti-mCherry Abs recognize the red-shifted fluorescent proteins (FPs) but not EGFP. HEK293 cells were transfected with DNA encoding various FPs, fixed and immunostained with biotinylated anti-mCherry clone 1 and visualized by Cy2- or Cy3-conjugated streptavidin. Fluorescence following transfection and immunostaining is shown in the left panel and middle panel, respectively. Merge images are shown in the right panel. (B) The specificity of the clones was examined by ELISA. Supernatants/cell media and cell lysates of transfected HEK293 were harvested and analysed in different dilutions (presented as log10 of dilution) by an ELISA. Clone 1 was used as coat and biotinylated clone 2 as detection antibody and vice versa. (C) The cell lysates analysed in B were also subjected to SDS-PAGE and Western blotting. The FPs were detected by anti-mCherry clone and chemiluminescence.

    Journal: Scandinavian Journal of Immunology

    Article Title: Generation of Antibody-Producing Hybridomas Following One Single Immunization with a Targeted DNA Vaccine

    doi: 10.1111/j.1365-3083.2011.02639.x

    Figure Lengend Snippet: Specificity of the anti-mCherry Abs clone 1 and 2. (A) Microscopy revealed that the anti-mCherry Abs recognize the red-shifted fluorescent proteins (FPs) but not EGFP. HEK293 cells were transfected with DNA encoding various FPs, fixed and immunostained with biotinylated anti-mCherry clone 1 and visualized by Cy2- or Cy3-conjugated streptavidin. Fluorescence following transfection and immunostaining is shown in the left panel and middle panel, respectively. Merge images are shown in the right panel. (B) The specificity of the clones was examined by ELISA. Supernatants/cell media and cell lysates of transfected HEK293 were harvested and analysed in different dilutions (presented as log10 of dilution) by an ELISA. Clone 1 was used as coat and biotinylated clone 2 as detection antibody and vice versa. (C) The cell lysates analysed in B were also subjected to SDS-PAGE and Western blotting. The FPs were detected by anti-mCherry clone and chemiluminescence.

    Article Snippet: Samples (50 μl/well) were incubated overnight followed by incubation with 75 μl/well of biotinylated detection antibodies [clone 1 or 2 (1 μg/ml) or class or subclass specific antibodies] (incubation time approximately 1.5 h) and alkaline phosphatase-conjugated streptavidin (1:3000, approximately 1.5 h) (GE Healthcare).

    Techniques: Microscopy, Transfection, Fluorescence, Immunostaining, Clone Assay, Enzyme-linked Immunosorbent Assay, SDS Page, Western Blot

    Targeting of mCherry towards major histocompatibility complex (MHC) class II enhanced the level of mCherry-specific Abs. (A) The targeting unit of [scFvI-E-mCherry] 2 binds specifically to MHC class II. Supernatants harvested from transfected HEK293 were used to stain EβdEαk- or Dd-transfected L cells. The bound vaccine proteins were detected by biotinylated anti-mCherry, streptavidin-PE and flow cytometry. (B) BALB/ c were given DNA encoding [scFvI-E-mCherry] 2 , [MIP1α-mCherry] 2 or mCherry i.m. before electroporation. Blood was harvested after 14 days, and the titres of mCherry-specific IgG was determined by ELISA ( n = 5 mice/group). (C) Time line. BALB/ c were immunized as described previously, and draining lymph nodes (LN) and spleen were harvested for generation of hybridomas. (D) ELISA measurements of mCherry-specific Abs following fusion of draining LN with OURI cells. DNA encoding [scFvI-E-mCherry] 2 or mCherry was injected into quadriceps of BALB/ c before electroporation ( n = 3 mice/group). After 29 days, the draining LN were harvested, fused with OURI cells and cultivated in 96-well plates. The cell supernatants were analysed in ELISA containing mCherry as coat.

    Journal: Scandinavian Journal of Immunology

    Article Title: Generation of Antibody-Producing Hybridomas Following One Single Immunization with a Targeted DNA Vaccine

    doi: 10.1111/j.1365-3083.2011.02639.x

    Figure Lengend Snippet: Targeting of mCherry towards major histocompatibility complex (MHC) class II enhanced the level of mCherry-specific Abs. (A) The targeting unit of [scFvI-E-mCherry] 2 binds specifically to MHC class II. Supernatants harvested from transfected HEK293 were used to stain EβdEαk- or Dd-transfected L cells. The bound vaccine proteins were detected by biotinylated anti-mCherry, streptavidin-PE and flow cytometry. (B) BALB/ c were given DNA encoding [scFvI-E-mCherry] 2 , [MIP1α-mCherry] 2 or mCherry i.m. before electroporation. Blood was harvested after 14 days, and the titres of mCherry-specific IgG was determined by ELISA ( n = 5 mice/group). (C) Time line. BALB/ c were immunized as described previously, and draining lymph nodes (LN) and spleen were harvested for generation of hybridomas. (D) ELISA measurements of mCherry-specific Abs following fusion of draining LN with OURI cells. DNA encoding [scFvI-E-mCherry] 2 or mCherry was injected into quadriceps of BALB/ c before electroporation ( n = 3 mice/group). After 29 days, the draining LN were harvested, fused with OURI cells and cultivated in 96-well plates. The cell supernatants were analysed in ELISA containing mCherry as coat.

    Article Snippet: Samples (50 μl/well) were incubated overnight followed by incubation with 75 μl/well of biotinylated detection antibodies [clone 1 or 2 (1 μg/ml) or class or subclass specific antibodies] (incubation time approximately 1.5 h) and alkaline phosphatase-conjugated streptavidin (1:3000, approximately 1.5 h) (GE Healthcare).

    Techniques: Transfection, Staining, Flow Cytometry, Cytometry, Electroporation, Enzyme-linked Immunosorbent Assay, Mouse Assay, Injection