histone h3ac antibody sampler kit  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc histone h3ac antibody sampler kit
    (A) GST pull down assay was performed to detect the binding of HIS-GST-tagged OscobB with recombinant histones H3 and H4 (NEB). Here, 12% SDS-PAGE gel shows OscobB can physically bind to both the histones H3 and H4. GST protein alone was used as a negative control in lanes 8 and 9 suggesting that the histones are not non-specifically binding with GST. (B) Detection of deacetylase activity of OscobB: Deacetylation assay of histone H3 from rice leaf nuclear extract was performed at 28°C. The reaction mixture containing OscobB was collected at different time points (0-105 mins). Sample reaction was stopped by boiling them with sample dye and then loading on a 12% SDS-PAGE gel. Western blot analysis was performed using <t>H3Ac</t> primary antibody to detect the extent of deacetylation by OscobB. Here the loading control is histone H3. (C) Detection of the deacetylation of specific lysine residues in histone H3 by OscobB. Lane 1- Empty pET28a vector, lane 2- purified recombinant OscobB, lane 3- purified HsSIRT6. Anti-H3 antibody was used as loading control for all the western blots. Empty vector and HsSIRT6 were used as negative and positive control, respectively. Coomassie brilliant blue gel shows the reaction mix containing purified OscobB along with histone H3 in lane 2 and purified HsSIRT6 along with histone H3 in lane 3. (D) Saturation kinetics for the OscobB enzyme activity: MM plot showing the OscobB deacetylation of H3K9Ac and H3K18Ac from rice leaf nuclear extract using varied concentrations of NAD + (0-600 μM). The non-linear regression plots were calculated using Graphpad Prism 8.3.0. The error bar depicts the S.D.; n=3. Further kinetic parameters ( K m and k cat values) were calculated using these plots.
    Histone H3ac Antibody Sampler Kit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histone h3ac antibody sampler kit/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    histone h3ac antibody sampler kit - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "A cobB like protein in Oryza sativa indica regulates the mitochondrial machinery under stress conditions"

    Article Title: A cobB like protein in Oryza sativa indica regulates the mitochondrial machinery under stress conditions

    Journal: bioRxiv

    doi: 10.1101/2022.04.17.488584

    (A) GST pull down assay was performed to detect the binding of HIS-GST-tagged OscobB with recombinant histones H3 and H4 (NEB). Here, 12% SDS-PAGE gel shows OscobB can physically bind to both the histones H3 and H4. GST protein alone was used as a negative control in lanes 8 and 9 suggesting that the histones are not non-specifically binding with GST. (B) Detection of deacetylase activity of OscobB: Deacetylation assay of histone H3 from rice leaf nuclear extract was performed at 28°C. The reaction mixture containing OscobB was collected at different time points (0-105 mins). Sample reaction was stopped by boiling them with sample dye and then loading on a 12% SDS-PAGE gel. Western blot analysis was performed using H3Ac primary antibody to detect the extent of deacetylation by OscobB. Here the loading control is histone H3. (C) Detection of the deacetylation of specific lysine residues in histone H3 by OscobB. Lane 1- Empty pET28a vector, lane 2- purified recombinant OscobB, lane 3- purified HsSIRT6. Anti-H3 antibody was used as loading control for all the western blots. Empty vector and HsSIRT6 were used as negative and positive control, respectively. Coomassie brilliant blue gel shows the reaction mix containing purified OscobB along with histone H3 in lane 2 and purified HsSIRT6 along with histone H3 in lane 3. (D) Saturation kinetics for the OscobB enzyme activity: MM plot showing the OscobB deacetylation of H3K9Ac and H3K18Ac from rice leaf nuclear extract using varied concentrations of NAD + (0-600 μM). The non-linear regression plots were calculated using Graphpad Prism 8.3.0. The error bar depicts the S.D.; n=3. Further kinetic parameters ( K m and k cat values) were calculated using these plots.
    Figure Legend Snippet: (A) GST pull down assay was performed to detect the binding of HIS-GST-tagged OscobB with recombinant histones H3 and H4 (NEB). Here, 12% SDS-PAGE gel shows OscobB can physically bind to both the histones H3 and H4. GST protein alone was used as a negative control in lanes 8 and 9 suggesting that the histones are not non-specifically binding with GST. (B) Detection of deacetylase activity of OscobB: Deacetylation assay of histone H3 from rice leaf nuclear extract was performed at 28°C. The reaction mixture containing OscobB was collected at different time points (0-105 mins). Sample reaction was stopped by boiling them with sample dye and then loading on a 12% SDS-PAGE gel. Western blot analysis was performed using H3Ac primary antibody to detect the extent of deacetylation by OscobB. Here the loading control is histone H3. (C) Detection of the deacetylation of specific lysine residues in histone H3 by OscobB. Lane 1- Empty pET28a vector, lane 2- purified recombinant OscobB, lane 3- purified HsSIRT6. Anti-H3 antibody was used as loading control for all the western blots. Empty vector and HsSIRT6 were used as negative and positive control, respectively. Coomassie brilliant blue gel shows the reaction mix containing purified OscobB along with histone H3 in lane 2 and purified HsSIRT6 along with histone H3 in lane 3. (D) Saturation kinetics for the OscobB enzyme activity: MM plot showing the OscobB deacetylation of H3K9Ac and H3K18Ac from rice leaf nuclear extract using varied concentrations of NAD + (0-600 μM). The non-linear regression plots were calculated using Graphpad Prism 8.3.0. The error bar depicts the S.D.; n=3. Further kinetic parameters ( K m and k cat values) were calculated using these plots.

    Techniques Used: Pull Down Assay, Binding Assay, Recombinant, SDS Page, Negative Control, Histone Deacetylase Assay, Activity Assay, Western Blot, Plasmid Preparation, Purification, Positive Control

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    Cell Signaling Technology Inc histone h3ac antibody sampler kit
    (A) GST pull down assay was performed to detect the binding of HIS-GST-tagged OscobB with recombinant histones H3 and H4 (NEB). Here, 12% SDS-PAGE gel shows OscobB can physically bind to both the histones H3 and H4. GST protein alone was used as a negative control in lanes 8 and 9 suggesting that the histones are not non-specifically binding with GST. (B) Detection of deacetylase activity of OscobB: Deacetylation assay of histone H3 from rice leaf nuclear extract was performed at 28°C. The reaction mixture containing OscobB was collected at different time points (0-105 mins). Sample reaction was stopped by boiling them with sample dye and then loading on a 12% SDS-PAGE gel. Western blot analysis was performed using <t>H3Ac</t> primary antibody to detect the extent of deacetylation by OscobB. Here the loading control is histone H3. (C) Detection of the deacetylation of specific lysine residues in histone H3 by OscobB. Lane 1- Empty pET28a vector, lane 2- purified recombinant OscobB, lane 3- purified HsSIRT6. Anti-H3 antibody was used as loading control for all the western blots. Empty vector and HsSIRT6 were used as negative and positive control, respectively. Coomassie brilliant blue gel shows the reaction mix containing purified OscobB along with histone H3 in lane 2 and purified HsSIRT6 along with histone H3 in lane 3. (D) Saturation kinetics for the OscobB enzyme activity: MM plot showing the OscobB deacetylation of H3K9Ac and H3K18Ac from rice leaf nuclear extract using varied concentrations of NAD + (0-600 μM). The non-linear regression plots were calculated using Graphpad Prism 8.3.0. The error bar depicts the S.D.; n=3. Further kinetic parameters ( K m and k cat values) were calculated using these plots.
    Histone H3ac Antibody Sampler Kit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/histone h3ac antibody sampler kit/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    histone h3ac antibody sampler kit - by Bioz Stars, 2023-03
    93/100 stars
    		  Buy from Supplier

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    (A) GST pull down assay was performed to detect the binding of HIS-GST-tagged OscobB with recombinant histones H3 and H4 (NEB). Here, 12% SDS-PAGE gel shows OscobB can physically bind to both the histones H3 and H4. GST protein alone was used as a negative control in lanes 8 and 9 suggesting that the histones are not non-specifically binding with GST. (B) Detection of deacetylase activity of OscobB: Deacetylation assay of histone H3 from rice leaf nuclear extract was performed at 28°C. The reaction mixture containing OscobB was collected at different time points (0-105 mins). Sample reaction was stopped by boiling them with sample dye and then loading on a 12% SDS-PAGE gel. Western blot analysis was performed using H3Ac primary antibody to detect the extent of deacetylation by OscobB. Here the loading control is histone H3. (C) Detection of the deacetylation of specific lysine residues in histone H3 by OscobB. Lane 1- Empty pET28a vector, lane 2- purified recombinant OscobB, lane 3- purified HsSIRT6. Anti-H3 antibody was used as loading control for all the western blots. Empty vector and HsSIRT6 were used as negative and positive control, respectively. Coomassie brilliant blue gel shows the reaction mix containing purified OscobB along with histone H3 in lane 2 and purified HsSIRT6 along with histone H3 in lane 3. (D) Saturation kinetics for the OscobB enzyme activity: MM plot showing the OscobB deacetylation of H3K9Ac and H3K18Ac from rice leaf nuclear extract using varied concentrations of NAD + (0-600 μM). The non-linear regression plots were calculated using Graphpad Prism 8.3.0. The error bar depicts the S.D.; n=3. Further kinetic parameters ( K m and k cat values) were calculated using these plots.

    Journal: bioRxiv

    Article Title: A cobB like protein in Oryza sativa indica regulates the mitochondrial machinery under stress conditions

    doi: 10.1101/2022.04.17.488584

    Figure Lengend Snippet: (A) GST pull down assay was performed to detect the binding of HIS-GST-tagged OscobB with recombinant histones H3 and H4 (NEB). Here, 12% SDS-PAGE gel shows OscobB can physically bind to both the histones H3 and H4. GST protein alone was used as a negative control in lanes 8 and 9 suggesting that the histones are not non-specifically binding with GST. (B) Detection of deacetylase activity of OscobB: Deacetylation assay of histone H3 from rice leaf nuclear extract was performed at 28°C. The reaction mixture containing OscobB was collected at different time points (0-105 mins). Sample reaction was stopped by boiling them with sample dye and then loading on a 12% SDS-PAGE gel. Western blot analysis was performed using H3Ac primary antibody to detect the extent of deacetylation by OscobB. Here the loading control is histone H3. (C) Detection of the deacetylation of specific lysine residues in histone H3 by OscobB. Lane 1- Empty pET28a vector, lane 2- purified recombinant OscobB, lane 3- purified HsSIRT6. Anti-H3 antibody was used as loading control for all the western blots. Empty vector and HsSIRT6 were used as negative and positive control, respectively. Coomassie brilliant blue gel shows the reaction mix containing purified OscobB along with histone H3 in lane 2 and purified HsSIRT6 along with histone H3 in lane 3. (D) Saturation kinetics for the OscobB enzyme activity: MM plot showing the OscobB deacetylation of H3K9Ac and H3K18Ac from rice leaf nuclear extract using varied concentrations of NAD + (0-600 μM). The non-linear regression plots were calculated using Graphpad Prism 8.3.0. The error bar depicts the S.D.; n=3. Further kinetic parameters ( K m and k cat values) were calculated using these plots.

    Article Snippet: The histone H3Ac antibody sampler kit (9927T) and acetylated Histone H4 antibody sampler kit (ab218056) used were purchased from Cell Signaling Technology (CST) and abcam (USA), respectively.

    Techniques: Pull Down Assay, Binding Assay, Recombinant, SDS Page, Negative Control, Histone Deacetylase Assay, Activity Assay, Western Blot, Plasmid Preparation, Purification, Positive Control