streptavidin horseradish peroxidase hrp reporter reagent  (Thermo Fisher)


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    Structured Review

    Thermo Fisher streptavidin horseradish peroxidase hrp reporter reagent
    Streptavidin Horseradish Peroxidase Hrp Reporter Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin horseradish peroxidase hrp reporter reagent/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin horseradish peroxidase hrp reporter reagent - by Bioz Stars, 2020-04
    91/100 stars

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    Labeling:

    Article Title: Rationally Designed Immunogens Targeting HIV-1 gp120 V1V2 Induce Distinct Conformation-Specific Antibody Responses in Rabbits
    Article Snippet: Plates were washed with PBS containing 0.02% Tween 20 before incubation (1 h at RT) with a streptavidin–horseradish peroxidase (HRP) reporter reagent (product 21130; Pierce). .. A reduction in signal reflects the presence of Abs in serum that competed with labeled MAbs for binding to the plate-bound antigen.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Rationally Designed Immunogens Targeting HIV-1 gp120 V1V2 Induce Distinct Conformation-Specific Antibody Responses in Rabbits
    Article Snippet: A competition ELISA was used to determine epitope specificity, i.e., whether sera from vaccinated animals contained Abs recognizing epitopes similar to the well-characterized Env-specific MAbs PG9, CH58, 697-D, and 830A. .. Plates were washed with PBS containing 0.02% Tween 20 before incubation (1 h at RT) with a streptavidin–horseradish peroxidase (HRP) reporter reagent (product 21130; Pierce).

    Concentration Assay:

    Article Title: Rationally Designed Immunogens Targeting HIV-1 gp120 V1V2 Induce Distinct Conformation-Specific Antibody Responses in Rabbits
    Article Snippet: The concentration for each biotinylated MAb used was predetermined in competition with its nonbiotinylated IgG. .. Plates were washed with PBS containing 0.02% Tween 20 before incubation (1 h at RT) with a streptavidin–horseradish peroxidase (HRP) reporter reagent (product 21130; Pierce).

    Incubation:

    Article Title: Rationally Designed Immunogens Targeting HIV-1 gp120 V1V2 Induce Distinct Conformation-Specific Antibody Responses in Rabbits
    Article Snippet: .. Plates were washed with PBS containing 0.02% Tween 20 before incubation (1 h at RT) with a streptavidin–horseradish peroxidase (HRP) reporter reagent (product 21130; Pierce). .. A reduction in signal reflects the presence of Abs in serum that competed with labeled MAbs for binding to the plate-bound antigen.

    Binding Assay:

    Article Title: Rationally Designed Immunogens Targeting HIV-1 gp120 V1V2 Induce Distinct Conformation-Specific Antibody Responses in Rabbits
    Article Snippet: Paragraph title: Competition antibody binding assay. ... Plates were washed with PBS containing 0.02% Tween 20 before incubation (1 h at RT) with a streptavidin–horseradish peroxidase (HRP) reporter reagent (product 21130; Pierce).

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    Thermo Fisher streptavidin hrp conjugate
    Target-selective protein labelling with LDNASA chemistry. a Western blotting analysis of FKBP12 labelling in cell lysates. HeLa cell lysate was mixed with recombinant FKBP12 (1 µM) and incubated with reagent in 50 mM HEPES buffer, pH 7.2, 37 °C, 1 h. <t>SAv-HRP,</t> <t>streptavidin–horseradish</t> peroxidase conjugate; CBB, Coomassie brilliant blue staining. The high background (smear) signals of lane 6 and 7 may suggest unspecific labelling due to a high concentration of reagent (20 µM), whereas such background signals were less in lanes 1–5 (1–5 µM of reagent). b Western blotting analysis of endogenous FKBP12 labelling in live C2C12 cells. The cells were treated with 1 or 5 (1 µM) for 0–120 min at 37 °C in medium. c Time course plot of the endogenous FKBP12 labelling with 1 . d Molecular structures of LDNASA 9 and LDAI 10 for FR labelling. e In-gel fluorescence and western blotting analysis of endogenous FR labelling in KB cells. The cells were treated with 9 or 10 (1 µM) for 0–60 min at 37 °C in medium. FA, Folic acid (competitive inhibitor). f Time profile of the endogenous FR labelling with 9 . I and I 30 min are band intensities of Oregon Green-labelled FR at the indicated time point and at 30 min, respectively. Error bars represent s.d., n = 3
    Streptavidin Hrp Conjugate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin hrp conjugate/product/Thermo Fisher
    Average 99 stars, based on 222 article reviews
    Price from $9.99 to $1999.99
    streptavidin hrp conjugate - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher streptavidin hrp
    Characterization of the BRWD2/PHIP CryptoTudor domain. ( A ) Diagram depicting the recombinant BRWD2/PHIP CryptoTudor–bromodomain (BRWD2/PHIP–Crypt–bromo) construct used for binding studies. N-terminal 10X-Histidine (red box) and C-terminal 1XFlag (green box) tags were included for protein purification. ( B ) Coomassie stained SDS-PAGE gel of in vitro chromatin capture experiment using the BRWD2/PHIP–Crypt–bromo module. (Lane 1 ) Input MNase-digested chromatin. (Lane 2 ) Chromatin subjected to anti-Flag immunoprecipitation without BRWD2/PHIP–Crypt–bromo. (Lane 3 ) Chromatin incubated with recombinant BRWD2/PHIP–Crypt–bromo and subjected to anti-Flag immunoprecipitation. ( C ) Quantitative MS analysis of histones captured in B . Bars indicate the fraction of total histone represented by each modification state in the input (gray) or Flag immunoprecipitation material (red). (Un) Unmodified; (me1) monomethylated; (me2) dimethylated; (me3) trimethylated; (Ac) acetylated. ( D ) Coomassie-stained SDS-PAGE gel of histone peptide pull-downs performed with human ( top ) or Drosophila ( bottom ) Crypt–bromo constructs. Recombinant protein was incubated with <t>streptavidin</t> beads alone (SA-beads) or the biotinylated histone peptides indicated. Ten percent of input and 20% of each pull-down sample were loaded. ( Bottom ) Eluted proteins were subjected to dot blotting using <t>streptavidin-HRP</t> (SA-HRP). ( E ) ITC experiments with human BRWD2–Crypt–bromo titrated with H3 unmodified ( left ), H3K4me1 ( middle ), and H3K4me3 ( right ) peptides. ( F ) Coomassie-stained SDS-PAGE gel of pull-downs performed with human BRWD2/PHIP–Crypt–bromo and a panel of histone peptides. Ten percent of input and 20% of each pull-down sample were loaded. Streptavidin-HRP dot blot loading control is shown below the gel image. ( G ) Coomassie-stained SDS-PAGE gel of a panel of histone peptide pull-downs performed with human BRWD2–Crypt–bromo ( top ), the isolated CryptoTudor domain ( middle ), or the isolated tandem bromodomains ( bottom ). Ten percent of input and 20% of each pull-down sample were loaded. Dot blot loading control is shown below each gel image. ( H ) Coomassie-stained gels of histone peptide pull-downs performed with point mutations of the BRWD2 CryptoTudor domain. Dot blot loading controls are shown below the gels. ( I ). Positions of residues predicted to be involved in methyl-lysine binding are highlighted in red.
    Streptavidin Hrp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 162 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin hrp/product/Thermo Fisher
    Average 99 stars, based on 162 article reviews
    Price from $9.99 to $1999.99
    streptavidin hrp - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher hrp conjugated streptavidin
    Characterization of ZASP-skeletal muscle α-actin 1 (ACTA1) interaction in mammalian cells and in vitro with purified proteins. A , co-IP ( IP ) assay showing the interaction between the ZASP isoforms and ACTA1 in wild-type mouse vastus muscle lysates. B and C , co-IP assays showing the interaction between the indicated ZASP isoforms and ACTA1 in non-muscle (COS7/HEK293) cells. The proteins were detected with anti-FLAG and anti-HA antibodies. D , purified GST-tagged ZASP-LΔex10 (WT, A165V, and A147T), but not GST alone, binds to biotinylated G-actin ( G-actin ) in a slot blot overlay assay. Biotin-G-actin was detected with <t>HRP-streptavidin.</t> GST and GST-ZASP were detected by immunoblotting with GST antibody ( anti-GST ). Arp2/3 and BSA served as positive and negative controls for G-actin binding. E , high-speed cosedimentation assay of purified GST or the indicated GST-ZASP-LΔex10 proteins and F-actin. Representative colloidal blue-stained gel images demonstrate a shift of the indicated GST-ZASP proteins from the supernatant ( S ) to the pellet ( P ) fraction in the presence of F-actin, whereas GST remained in the supernatant. Nonlinear regression analysis of the ZASP-actin interaction is shown in the right panel (triplicate assays, data represent mean ± S.D.).
    Hrp Conjugated Streptavidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 183 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated streptavidin/product/Thermo Fisher
    Average 99 stars, based on 183 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated streptavidin - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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    Target-selective protein labelling with LDNASA chemistry. a Western blotting analysis of FKBP12 labelling in cell lysates. HeLa cell lysate was mixed with recombinant FKBP12 (1 µM) and incubated with reagent in 50 mM HEPES buffer, pH 7.2, 37 °C, 1 h. SAv-HRP, streptavidin–horseradish peroxidase conjugate; CBB, Coomassie brilliant blue staining. The high background (smear) signals of lane 6 and 7 may suggest unspecific labelling due to a high concentration of reagent (20 µM), whereas such background signals were less in lanes 1–5 (1–5 µM of reagent). b Western blotting analysis of endogenous FKBP12 labelling in live C2C12 cells. The cells were treated with 1 or 5 (1 µM) for 0–120 min at 37 °C in medium. c Time course plot of the endogenous FKBP12 labelling with 1 . d Molecular structures of LDNASA 9 and LDAI 10 for FR labelling. e In-gel fluorescence and western blotting analysis of endogenous FR labelling in KB cells. The cells were treated with 9 or 10 (1 µM) for 0–60 min at 37 °C in medium. FA, Folic acid (competitive inhibitor). f Time profile of the endogenous FR labelling with 9 . I and I 30 min are band intensities of Oregon Green-labelled FR at the indicated time point and at 30 min, respectively. Error bars represent s.d., n = 3

    Journal: Nature Communications

    Article Title: Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide

    doi: 10.1038/s41467-018-04343-0

    Figure Lengend Snippet: Target-selective protein labelling with LDNASA chemistry. a Western blotting analysis of FKBP12 labelling in cell lysates. HeLa cell lysate was mixed with recombinant FKBP12 (1 µM) and incubated with reagent in 50 mM HEPES buffer, pH 7.2, 37 °C, 1 h. SAv-HRP, streptavidin–horseradish peroxidase conjugate; CBB, Coomassie brilliant blue staining. The high background (smear) signals of lane 6 and 7 may suggest unspecific labelling due to a high concentration of reagent (20 µM), whereas such background signals were less in lanes 1–5 (1–5 µM of reagent). b Western blotting analysis of endogenous FKBP12 labelling in live C2C12 cells. The cells were treated with 1 or 5 (1 µM) for 0–120 min at 37 °C in medium. c Time course plot of the endogenous FKBP12 labelling with 1 . d Molecular structures of LDNASA 9 and LDAI 10 for FR labelling. e In-gel fluorescence and western blotting analysis of endogenous FR labelling in KB cells. The cells were treated with 9 or 10 (1 µM) for 0–60 min at 37 °C in medium. FA, Folic acid (competitive inhibitor). f Time profile of the endogenous FR labelling with 9 . I and I 30 min are band intensities of Oregon Green-labelled FR at the indicated time point and at 30 min, respectively. Error bars represent s.d., n = 3

    Article Snippet: The samples were analysed by western blotting using Streptavidin-HRP conjugate (SAv-HRP, Thermo, S911, 1:5000) and Coomassie Brilliant Blue (CBB) stain.

    Techniques: Western Blot, Recombinant, Incubation, Staining, Concentration Assay, Fluorescence

    Restoring protein synthesis delays retinal degeneration. (A) Protein synthesis levels depicted by the detection of azidohomoalanine incorporation into proteins with HRP-conjugated streptavidin and (B) coomassie staining for normalization of the membrane. (C) Graph showing relative levels of protein synthesis in the retinas of C57BL/6J (n = 3), rd16 (n = 3), and rd16 4ebp1/2 −/− (n = 3) mice at P15. (D) ERG waveforms of three groups and (E) graph showing mean ERG a- and b- wave amplitudes of C57BL/6J (n = 4), rd16 (n = 6), and rd16 4ebp1/2 −/− (n = 6) mice at P17. (F) Representative images of H E-stained retinal sections taken at P18 in the regions denoted as * in (G). (G) Spider plots depicting number of photoreceptor nuclei in the ONL as counted by a masked investigator. The spidergram was generated by plotting the number of nuclei using 200 μm step in the distance from the ONH for both hemispheres. (H) TUNEL staining of retinal sections from C57BL/6J (n = 3), rd16 (n = 4), and rd16 4ebp1/2 −/− (n = 3) eyes at P15. (I) Graph showing TUNEL-positive nuclei per square millimeter in the three groups. Green, TUNEL; blue, DAPI; a.u, arbitrary units; kD, kilodaltons. Error bars: SEM. Statistical significance denoted by *P

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Role of Translational Attenuation in Inherited Retinal Degeneration

    doi: 10.1167/iovs.19-27512

    Figure Lengend Snippet: Restoring protein synthesis delays retinal degeneration. (A) Protein synthesis levels depicted by the detection of azidohomoalanine incorporation into proteins with HRP-conjugated streptavidin and (B) coomassie staining for normalization of the membrane. (C) Graph showing relative levels of protein synthesis in the retinas of C57BL/6J (n = 3), rd16 (n = 3), and rd16 4ebp1/2 −/− (n = 3) mice at P15. (D) ERG waveforms of three groups and (E) graph showing mean ERG a- and b- wave amplitudes of C57BL/6J (n = 4), rd16 (n = 6), and rd16 4ebp1/2 −/− (n = 6) mice at P17. (F) Representative images of H E-stained retinal sections taken at P18 in the regions denoted as * in (G). (G) Spider plots depicting number of photoreceptor nuclei in the ONL as counted by a masked investigator. The spidergram was generated by plotting the number of nuclei using 200 μm step in the distance from the ONH for both hemispheres. (H) TUNEL staining of retinal sections from C57BL/6J (n = 3), rd16 (n = 4), and rd16 4ebp1/2 −/− (n = 3) eyes at P15. (I) Graph showing TUNEL-positive nuclei per square millimeter in the three groups. Green, TUNEL; blue, DAPI; a.u, arbitrary units; kD, kilodaltons. Error bars: SEM. Statistical significance denoted by *P

    Article Snippet: Biotinylated proteins then were probed with HRP-conjugated streptavidin (S911; Thermo Fisher Scientific), incubated in ECL substrate (RPN2232; GE Healthcare, Chicago, IL, USA) and then imaged using a LI-COR (Lincoln, NE, USA) Fc imaging system.

    Techniques: Staining, Mouse Assay, Generated, TUNEL Assay

    Characterization of the BRWD2/PHIP CryptoTudor domain. ( A ) Diagram depicting the recombinant BRWD2/PHIP CryptoTudor–bromodomain (BRWD2/PHIP–Crypt–bromo) construct used for binding studies. N-terminal 10X-Histidine (red box) and C-terminal 1XFlag (green box) tags were included for protein purification. ( B ) Coomassie stained SDS-PAGE gel of in vitro chromatin capture experiment using the BRWD2/PHIP–Crypt–bromo module. (Lane 1 ) Input MNase-digested chromatin. (Lane 2 ) Chromatin subjected to anti-Flag immunoprecipitation without BRWD2/PHIP–Crypt–bromo. (Lane 3 ) Chromatin incubated with recombinant BRWD2/PHIP–Crypt–bromo and subjected to anti-Flag immunoprecipitation. ( C ) Quantitative MS analysis of histones captured in B . Bars indicate the fraction of total histone represented by each modification state in the input (gray) or Flag immunoprecipitation material (red). (Un) Unmodified; (me1) monomethylated; (me2) dimethylated; (me3) trimethylated; (Ac) acetylated. ( D ) Coomassie-stained SDS-PAGE gel of histone peptide pull-downs performed with human ( top ) or Drosophila ( bottom ) Crypt–bromo constructs. Recombinant protein was incubated with streptavidin beads alone (SA-beads) or the biotinylated histone peptides indicated. Ten percent of input and 20% of each pull-down sample were loaded. ( Bottom ) Eluted proteins were subjected to dot blotting using streptavidin-HRP (SA-HRP). ( E ) ITC experiments with human BRWD2–Crypt–bromo titrated with H3 unmodified ( left ), H3K4me1 ( middle ), and H3K4me3 ( right ) peptides. ( F ) Coomassie-stained SDS-PAGE gel of pull-downs performed with human BRWD2/PHIP–Crypt–bromo and a panel of histone peptides. Ten percent of input and 20% of each pull-down sample were loaded. Streptavidin-HRP dot blot loading control is shown below the gel image. ( G ) Coomassie-stained SDS-PAGE gel of a panel of histone peptide pull-downs performed with human BRWD2–Crypt–bromo ( top ), the isolated CryptoTudor domain ( middle ), or the isolated tandem bromodomains ( bottom ). Ten percent of input and 20% of each pull-down sample were loaded. Dot blot loading control is shown below each gel image. ( H ) Coomassie-stained gels of histone peptide pull-downs performed with point mutations of the BRWD2 CryptoTudor domain. Dot blot loading controls are shown below the gels. ( I ). Positions of residues predicted to be involved in methyl-lysine binding are highlighted in red.

    Journal: Genes & Development

    Article Title: A cryptic Tudor domain links BRWD2/PHIP to COMPASS-mediated histone H3K4 methylation

    doi: 10.1101/gad.305201.117

    Figure Lengend Snippet: Characterization of the BRWD2/PHIP CryptoTudor domain. ( A ) Diagram depicting the recombinant BRWD2/PHIP CryptoTudor–bromodomain (BRWD2/PHIP–Crypt–bromo) construct used for binding studies. N-terminal 10X-Histidine (red box) and C-terminal 1XFlag (green box) tags were included for protein purification. ( B ) Coomassie stained SDS-PAGE gel of in vitro chromatin capture experiment using the BRWD2/PHIP–Crypt–bromo module. (Lane 1 ) Input MNase-digested chromatin. (Lane 2 ) Chromatin subjected to anti-Flag immunoprecipitation without BRWD2/PHIP–Crypt–bromo. (Lane 3 ) Chromatin incubated with recombinant BRWD2/PHIP–Crypt–bromo and subjected to anti-Flag immunoprecipitation. ( C ) Quantitative MS analysis of histones captured in B . Bars indicate the fraction of total histone represented by each modification state in the input (gray) or Flag immunoprecipitation material (red). (Un) Unmodified; (me1) monomethylated; (me2) dimethylated; (me3) trimethylated; (Ac) acetylated. ( D ) Coomassie-stained SDS-PAGE gel of histone peptide pull-downs performed with human ( top ) or Drosophila ( bottom ) Crypt–bromo constructs. Recombinant protein was incubated with streptavidin beads alone (SA-beads) or the biotinylated histone peptides indicated. Ten percent of input and 20% of each pull-down sample were loaded. ( Bottom ) Eluted proteins were subjected to dot blotting using streptavidin-HRP (SA-HRP). ( E ) ITC experiments with human BRWD2–Crypt–bromo titrated with H3 unmodified ( left ), H3K4me1 ( middle ), and H3K4me3 ( right ) peptides. ( F ) Coomassie-stained SDS-PAGE gel of pull-downs performed with human BRWD2/PHIP–Crypt–bromo and a panel of histone peptides. Ten percent of input and 20% of each pull-down sample were loaded. Streptavidin-HRP dot blot loading control is shown below the gel image. ( G ) Coomassie-stained SDS-PAGE gel of a panel of histone peptide pull-downs performed with human BRWD2–Crypt–bromo ( top ), the isolated CryptoTudor domain ( middle ), or the isolated tandem bromodomains ( bottom ). Ten percent of input and 20% of each pull-down sample were loaded. Dot blot loading control is shown below each gel image. ( H ) Coomassie-stained gels of histone peptide pull-downs performed with point mutations of the BRWD2 CryptoTudor domain. Dot blot loading controls are shown below the gels. ( I ). Positions of residues predicted to be involved in methyl-lysine binding are highlighted in red.

    Article Snippet: As a loading control, 0.25 µL of the eluted proteins was spotted onto a nitrocellulose membrane and detected using streptavidin-HRP (Thermo Scientific, no. 21130).

    Techniques: Recombinant, Construct, Binding Assay, Protein Purification, Staining, SDS Page, In Vitro, Immunoprecipitation, Incubation, Mass Spectrometry, Modification, Dot Blot, Isolation

    Characterization of ZASP-skeletal muscle α-actin 1 (ACTA1) interaction in mammalian cells and in vitro with purified proteins. A , co-IP ( IP ) assay showing the interaction between the ZASP isoforms and ACTA1 in wild-type mouse vastus muscle lysates. B and C , co-IP assays showing the interaction between the indicated ZASP isoforms and ACTA1 in non-muscle (COS7/HEK293) cells. The proteins were detected with anti-FLAG and anti-HA antibodies. D , purified GST-tagged ZASP-LΔex10 (WT, A165V, and A147T), but not GST alone, binds to biotinylated G-actin ( G-actin ) in a slot blot overlay assay. Biotin-G-actin was detected with HRP-streptavidin. GST and GST-ZASP were detected by immunoblotting with GST antibody ( anti-GST ). Arp2/3 and BSA served as positive and negative controls for G-actin binding. E , high-speed cosedimentation assay of purified GST or the indicated GST-ZASP-LΔex10 proteins and F-actin. Representative colloidal blue-stained gel images demonstrate a shift of the indicated GST-ZASP proteins from the supernatant ( S ) to the pellet ( P ) fraction in the presence of F-actin, whereas GST remained in the supernatant. Nonlinear regression analysis of the ZASP-actin interaction is shown in the right panel (triplicate assays, data represent mean ± S.D.).

    Journal: The Journal of Biological Chemistry

    Article Title: Z-disc-associated, Alternatively Spliced, PDZ Motif-containing Protein (ZASP) Mutations in the Actin-binding Domain Cause Disruption of Skeletal Muscle Actin Filaments in Myofibrillar Myopathy *

    doi: 10.1074/jbc.M114.550418

    Figure Lengend Snippet: Characterization of ZASP-skeletal muscle α-actin 1 (ACTA1) interaction in mammalian cells and in vitro with purified proteins. A , co-IP ( IP ) assay showing the interaction between the ZASP isoforms and ACTA1 in wild-type mouse vastus muscle lysates. B and C , co-IP assays showing the interaction between the indicated ZASP isoforms and ACTA1 in non-muscle (COS7/HEK293) cells. The proteins were detected with anti-FLAG and anti-HA antibodies. D , purified GST-tagged ZASP-LΔex10 (WT, A165V, and A147T), but not GST alone, binds to biotinylated G-actin ( G-actin ) in a slot blot overlay assay. Biotin-G-actin was detected with HRP-streptavidin. GST and GST-ZASP were detected by immunoblotting with GST antibody ( anti-GST ). Arp2/3 and BSA served as positive and negative controls for G-actin binding. E , high-speed cosedimentation assay of purified GST or the indicated GST-ZASP-LΔex10 proteins and F-actin. Representative colloidal blue-stained gel images demonstrate a shift of the indicated GST-ZASP proteins from the supernatant ( S ) to the pellet ( P ) fraction in the presence of F-actin, whereas GST remained in the supernatant. Nonlinear regression analysis of the ZASP-actin interaction is shown in the right panel (triplicate assays, data represent mean ± S.D.).

    Article Snippet: The membrane was incubated with HRP-conjugated streptavidin (N100, Thermo Scientific), and G-actin binding was detected using chemiluminescence.

    Techniques: In Vitro, Purification, Co-Immunoprecipitation Assay, Dot Blot, Overlay Assay, Binding Assay, Staining