Structured Review

Millipore anti flag m2 magnetic beads
(A) Western blot of lysates of non-transfected HEK293T cells (-) or transiently transfected with constructs to express either BirA-Myc alone (empty), BirA-Myc-fused eGFP, or BirA-Myc-fused YTHDF2 or a version without its N-terminus (ΔNT). Biotinylation was analysed using Alexa488-labelled streptavidin, and expression levels of the fusion proteins were analyzed with antibodies against the Myc tag. GAPDH was used as a loading control. (B) Volcano plot showing proteins enriched in the YTHDF2 BioID over the control BioID samples (BirA-Myc alone + BirA-Myc-fused eGFP). The logarithmic fold-changes were plotted against negative logarithmic P values of a two-sided two samples t-test. A selection of silencing factors (red) and translation factors, including ribosomal proteins (blue) is indicated. The YTHDF paralogs are shown in green. (C) Volcano plot showing of the interactome of a version of YTHDF2 deleted of its N-terminus (region 1-389). The plot represents the differential enrichment of proteins detected in the BirA-Myc-YTHDF2 ΔNT dataset (positive values) calculated against data obtained with the negative controls, BirA-Myc alone and BirA-Myc-eGFP (negative values). Proteins involved in RNA splicing are shown in orange. (D) Interaction of YTHDF2 with its high confidence preys. Vectors expressing 3xFlag-tagged YTHDF2, either Full-Length (FL) or ΔNT, or 3xFlag as control (-), were transfected into HEK293T cells. Extracts were immunoprecipitated with <t>anti-Flag</t> antibody. Total lysates (input) and IP extracts were analyzed by Western blot (WB) with the indicated antibodies. (E) Heatmap showing the logarithmic fold-changes of the ribosomal proteins detected in the BioID datasets of BirA-Myc-fused YTHDF1, 2 or 3 over the control BioID samples (BirA-Myc alone and BirA-Myc-fused eGFP). Note that the white color indicate that the protein was not present in the MS dataset. (F) Ribo Mega-SEC fractionation of a HEK293T cell lysate. Extract was fractionated using a flow rate of 0.2 ml/min on a 2,000 Å SEC column. The chromatogram is shown in Supplementary Figure S2E. Western blot was performed with the indicated antibodies.
Anti Flag M2 Magnetic Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti flag m2 magnetic beads - by Bioz Stars, 2023-11
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1) Product Images from "The YTHDF1-3 proteins are bidirectionally influenced by the codon content of their mRNA targets"

Article Title: The YTHDF1-3 proteins are bidirectionally influenced by the codon content of their mRNA targets

Journal: bioRxiv

doi: 10.1101/2023.11.20.565808

(A) Western blot of lysates of non-transfected HEK293T cells (-) or transiently transfected with constructs to express either BirA-Myc alone (empty), BirA-Myc-fused eGFP, or BirA-Myc-fused YTHDF2 or a version without its N-terminus (ΔNT). Biotinylation was analysed using Alexa488-labelled streptavidin, and expression levels of the fusion proteins were analyzed with antibodies against the Myc tag. GAPDH was used as a loading control. (B) Volcano plot showing proteins enriched in the YTHDF2 BioID over the control BioID samples (BirA-Myc alone + BirA-Myc-fused eGFP). The logarithmic fold-changes were plotted against negative logarithmic P values of a two-sided two samples t-test. A selection of silencing factors (red) and translation factors, including ribosomal proteins (blue) is indicated. The YTHDF paralogs are shown in green. (C) Volcano plot showing of the interactome of a version of YTHDF2 deleted of its N-terminus (region 1-389). The plot represents the differential enrichment of proteins detected in the BirA-Myc-YTHDF2 ΔNT dataset (positive values) calculated against data obtained with the negative controls, BirA-Myc alone and BirA-Myc-eGFP (negative values). Proteins involved in RNA splicing are shown in orange. (D) Interaction of YTHDF2 with its high confidence preys. Vectors expressing 3xFlag-tagged YTHDF2, either Full-Length (FL) or ΔNT, or 3xFlag as control (-), were transfected into HEK293T cells. Extracts were immunoprecipitated with anti-Flag antibody. Total lysates (input) and IP extracts were analyzed by Western blot (WB) with the indicated antibodies. (E) Heatmap showing the logarithmic fold-changes of the ribosomal proteins detected in the BioID datasets of BirA-Myc-fused YTHDF1, 2 or 3 over the control BioID samples (BirA-Myc alone and BirA-Myc-fused eGFP). Note that the white color indicate that the protein was not present in the MS dataset. (F) Ribo Mega-SEC fractionation of a HEK293T cell lysate. Extract was fractionated using a flow rate of 0.2 ml/min on a 2,000 Å SEC column. The chromatogram is shown in Supplementary Figure S2E. Western blot was performed with the indicated antibodies.
Figure Legend Snippet: (A) Western blot of lysates of non-transfected HEK293T cells (-) or transiently transfected with constructs to express either BirA-Myc alone (empty), BirA-Myc-fused eGFP, or BirA-Myc-fused YTHDF2 or a version without its N-terminus (ΔNT). Biotinylation was analysed using Alexa488-labelled streptavidin, and expression levels of the fusion proteins were analyzed with antibodies against the Myc tag. GAPDH was used as a loading control. (B) Volcano plot showing proteins enriched in the YTHDF2 BioID over the control BioID samples (BirA-Myc alone + BirA-Myc-fused eGFP). The logarithmic fold-changes were plotted against negative logarithmic P values of a two-sided two samples t-test. A selection of silencing factors (red) and translation factors, including ribosomal proteins (blue) is indicated. The YTHDF paralogs are shown in green. (C) Volcano plot showing of the interactome of a version of YTHDF2 deleted of its N-terminus (region 1-389). The plot represents the differential enrichment of proteins detected in the BirA-Myc-YTHDF2 ΔNT dataset (positive values) calculated against data obtained with the negative controls, BirA-Myc alone and BirA-Myc-eGFP (negative values). Proteins involved in RNA splicing are shown in orange. (D) Interaction of YTHDF2 with its high confidence preys. Vectors expressing 3xFlag-tagged YTHDF2, either Full-Length (FL) or ΔNT, or 3xFlag as control (-), were transfected into HEK293T cells. Extracts were immunoprecipitated with anti-Flag antibody. Total lysates (input) and IP extracts were analyzed by Western blot (WB) with the indicated antibodies. (E) Heatmap showing the logarithmic fold-changes of the ribosomal proteins detected in the BioID datasets of BirA-Myc-fused YTHDF1, 2 or 3 over the control BioID samples (BirA-Myc alone and BirA-Myc-fused eGFP). Note that the white color indicate that the protein was not present in the MS dataset. (F) Ribo Mega-SEC fractionation of a HEK293T cell lysate. Extract was fractionated using a flow rate of 0.2 ml/min on a 2,000 Å SEC column. The chromatogram is shown in Supplementary Figure S2E. Western blot was performed with the indicated antibodies.

Techniques Used: Western Blot, Transfection, Construct, Expressing, Selection, Immunoprecipitation, Fractionation


Structured Review

Millipore mouse monoclonal anti flag m2 antibody
Mouse Monoclonal Anti Flag M2 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal anti flag m2 antibody/product/Millipore
Average 86 stars, based on 1 article reviews
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mouse monoclonal anti flag m2 antibody - by Bioz Stars, 2023-11
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Structured Review

Millipore mouse monoclonal anti flag m2
Mouse Monoclonal Anti Flag M2, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal anti flag m2/product/Millipore
Average 86 stars, based on 1 article reviews
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mouse monoclonal anti flag m2 - by Bioz Stars, 2023-11
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Structured Review

Millipore mouse monoclonal anti flag m2 antibody
Mouse Monoclonal Anti Flag M2 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal anti flag m2 antibody/product/Millipore
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
mouse monoclonal anti flag m2 antibody - by Bioz Stars, 2023-11
86/100 stars

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Millipore anti flag m2 affinity gel
Anti Flag M2 Affinity Gel, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti flag m2 affinity gel/product/Millipore
Average 86 stars, based on 1 article reviews
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anti flag m2 affinity gel - by Bioz Stars, 2023-11
86/100 stars

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Structured Review

Millipore anti flag m2 magnetic beads
Anti Flag M2 Magnetic Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti flag m2 magnetic beads/product/Millipore
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti flag m2 magnetic beads - by Bioz Stars, 2023-11
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Millipore anti flag m2 affinity gel beads
( A ) In vitro protein phosphatase (PP) assay. Representative immunoblots of p-NCC and t-NCC are shown for assays of NCC with PP1A (lanes 1 and 2), PP2A (lanes 5 and 6), and calcineurin A (CALNA) or vehicle (lanes 3, 7, and 11). For these studies, NCC was isolated by IP with an anti-NCC antibody (lanes 1, 2, 3, 5, 6, and 7) and compared with the negative control IgG (lanes 4, 8, and 12). ( B ) PP1A preferentially interacted with NCC in glutathione-agarose affinity chromatography assays with the GST fusion protein of the NCC terminus GST-NCC-(N), but not the negative control GST alone. Shown are representative immunoblot binding assays with recombinant PP1A, PP2A, and CALNA. Input protein phosphatase (lane 1) is shown relative to GST-bound (lane 2) or GST-NCC-(N) (lane 3). ( C ) A greater amount of PP1A co-immunoprecipitated with NCC when the extracellular K + concentration was high. Representative immunoblots show NCC, PP1A, and p-NCC (T58) in <t>anti-FLAG</t> immunoprecipitated samples relative to input from FLAG-tagged NCC-expressing MDCKI cells incubated with 3.5 mM (control), 0.5 mM (low), or 8 mM (high) K + buffers. Low-chloride buffer was used as a positive control to elevate p-NCC (T58) levels. Graphs show semiquantitative assessment of NCC, PP1, and p-NCC (T58) in NCC immunoprecipitated samples (bottom, right) relative to input. Data are from 3 individual experiments with 3 replicates for each condition ( n = 9). * P < 0.05, relative to the low-K + condition, by 1-way ANOVA followed by multiple-comparison test. Data are presented as the mean ± SEM. CALNA, calcineurin.
Anti Flag M2 Affinity Gel Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti flag m2 affinity gel beads/product/Millipore
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti flag m2 affinity gel beads - by Bioz Stars, 2023-11
86/100 stars

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1) Product Images from "Dietary potassium stimulates Ppp1Ca-Ppp1r1a dephosphorylation of kidney NaCl cotransporter and reduces blood pressure"

Article Title: Dietary potassium stimulates Ppp1Ca-Ppp1r1a dephosphorylation of kidney NaCl cotransporter and reduces blood pressure

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI158498

( A ) In vitro protein phosphatase (PP) assay. Representative immunoblots of p-NCC and t-NCC are shown for assays of NCC with PP1A (lanes 1 and 2), PP2A (lanes 5 and 6), and calcineurin A (CALNA) or vehicle (lanes 3, 7, and 11). For these studies, NCC was isolated by IP with an anti-NCC antibody (lanes 1, 2, 3, 5, 6, and 7) and compared with the negative control IgG (lanes 4, 8, and 12). ( B ) PP1A preferentially interacted with NCC in glutathione-agarose affinity chromatography assays with the GST fusion protein of the NCC terminus GST-NCC-(N), but not the negative control GST alone. Shown are representative immunoblot binding assays with recombinant PP1A, PP2A, and CALNA. Input protein phosphatase (lane 1) is shown relative to GST-bound (lane 2) or GST-NCC-(N) (lane 3). ( C ) A greater amount of PP1A co-immunoprecipitated with NCC when the extracellular K + concentration was high. Representative immunoblots show NCC, PP1A, and p-NCC (T58) in anti-FLAG immunoprecipitated samples relative to input from FLAG-tagged NCC-expressing MDCKI cells incubated with 3.5 mM (control), 0.5 mM (low), or 8 mM (high) K + buffers. Low-chloride buffer was used as a positive control to elevate p-NCC (T58) levels. Graphs show semiquantitative assessment of NCC, PP1, and p-NCC (T58) in NCC immunoprecipitated samples (bottom, right) relative to input. Data are from 3 individual experiments with 3 replicates for each condition ( n = 9). * P < 0.05, relative to the low-K + condition, by 1-way ANOVA followed by multiple-comparison test. Data are presented as the mean ± SEM. CALNA, calcineurin.
Figure Legend Snippet: ( A ) In vitro protein phosphatase (PP) assay. Representative immunoblots of p-NCC and t-NCC are shown for assays of NCC with PP1A (lanes 1 and 2), PP2A (lanes 5 and 6), and calcineurin A (CALNA) or vehicle (lanes 3, 7, and 11). For these studies, NCC was isolated by IP with an anti-NCC antibody (lanes 1, 2, 3, 5, 6, and 7) and compared with the negative control IgG (lanes 4, 8, and 12). ( B ) PP1A preferentially interacted with NCC in glutathione-agarose affinity chromatography assays with the GST fusion protein of the NCC terminus GST-NCC-(N), but not the negative control GST alone. Shown are representative immunoblot binding assays with recombinant PP1A, PP2A, and CALNA. Input protein phosphatase (lane 1) is shown relative to GST-bound (lane 2) or GST-NCC-(N) (lane 3). ( C ) A greater amount of PP1A co-immunoprecipitated with NCC when the extracellular K + concentration was high. Representative immunoblots show NCC, PP1A, and p-NCC (T58) in anti-FLAG immunoprecipitated samples relative to input from FLAG-tagged NCC-expressing MDCKI cells incubated with 3.5 mM (control), 0.5 mM (low), or 8 mM (high) K + buffers. Low-chloride buffer was used as a positive control to elevate p-NCC (T58) levels. Graphs show semiquantitative assessment of NCC, PP1, and p-NCC (T58) in NCC immunoprecipitated samples (bottom, right) relative to input. Data are from 3 individual experiments with 3 replicates for each condition ( n = 9). * P < 0.05, relative to the low-K + condition, by 1-way ANOVA followed by multiple-comparison test. Data are presented as the mean ± SEM. CALNA, calcineurin.

Techniques Used: In Vitro, Western Blot, Isolation, Negative Control, Affinity Chromatography, Binding Assay, Recombinant, Immunoprecipitation, Concentration Assay, Expressing, Incubation, Positive Control, Comparison


Structured Review

Millipore anti flag m2 affinity gel beads
( A ) In vitro protein phosphatase (PP) assay. Representative immunoblots of p-NCC and t-NCC are shown for assays of NCC with PP1A (lanes 1 and 2), PP2A (lanes 5 and 6), and calcineurin A (CALNA) or vehicle (lanes 3, 7, and 11). For these studies, NCC was isolated by IP with an anti-NCC antibody (lanes 1, 2, 3, 5, 6, and 7) and compared with the negative control IgG (lanes 4, 8, and 12). ( B ) PP1A preferentially interacted with NCC in glutathione-agarose affinity chromatography assays with the GST fusion protein of the NCC terminus GST-NCC-(N), but not the negative control GST alone. Shown are representative immunoblot binding assays with recombinant PP1A, PP2A, and CALNA. Input protein phosphatase (lane 1) is shown relative to GST-bound (lane 2) or GST-NCC-(N) (lane 3). ( C ) A greater amount of PP1A co-immunoprecipitated with NCC when the extracellular K + concentration was high. Representative immunoblots show NCC, PP1A, and p-NCC (T58) in <t>anti-FLAG</t> immunoprecipitated samples relative to input from FLAG-tagged NCC-expressing MDCKI cells incubated with 3.5 mM (control), 0.5 mM (low), or 8 mM (high) K + buffers. Low-chloride buffer was used as a positive control to elevate p-NCC (T58) levels. Graphs show semiquantitative assessment of NCC, PP1, and p-NCC (T58) in NCC immunoprecipitated samples (bottom, right) relative to input. Data are from 3 individual experiments with 3 replicates for each condition ( n = 9). * P < 0.05, relative to the low-K + condition, by 1-way ANOVA followed by multiple-comparison test. Data are presented as the mean ± SEM. CALNA, calcineurin.
Anti Flag M2 Affinity Gel Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti flag m2 affinity gel beads/product/Millipore
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti flag m2 affinity gel beads - by Bioz Stars, 2023-11
86/100 stars

Images

1) Product Images from "Dietary potassium stimulates Ppp1Ca-Ppp1r1a dephosphorylation of kidney NaCl cotransporter and reduces blood pressure"

Article Title: Dietary potassium stimulates Ppp1Ca-Ppp1r1a dephosphorylation of kidney NaCl cotransporter and reduces blood pressure

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI158498

( A ) In vitro protein phosphatase (PP) assay. Representative immunoblots of p-NCC and t-NCC are shown for assays of NCC with PP1A (lanes 1 and 2), PP2A (lanes 5 and 6), and calcineurin A (CALNA) or vehicle (lanes 3, 7, and 11). For these studies, NCC was isolated by IP with an anti-NCC antibody (lanes 1, 2, 3, 5, 6, and 7) and compared with the negative control IgG (lanes 4, 8, and 12). ( B ) PP1A preferentially interacted with NCC in glutathione-agarose affinity chromatography assays with the GST fusion protein of the NCC terminus GST-NCC-(N), but not the negative control GST alone. Shown are representative immunoblot binding assays with recombinant PP1A, PP2A, and CALNA. Input protein phosphatase (lane 1) is shown relative to GST-bound (lane 2) or GST-NCC-(N) (lane 3). ( C ) A greater amount of PP1A co-immunoprecipitated with NCC when the extracellular K + concentration was high. Representative immunoblots show NCC, PP1A, and p-NCC (T58) in anti-FLAG immunoprecipitated samples relative to input from FLAG-tagged NCC-expressing MDCKI cells incubated with 3.5 mM (control), 0.5 mM (low), or 8 mM (high) K + buffers. Low-chloride buffer was used as a positive control to elevate p-NCC (T58) levels. Graphs show semiquantitative assessment of NCC, PP1, and p-NCC (T58) in NCC immunoprecipitated samples (bottom, right) relative to input. Data are from 3 individual experiments with 3 replicates for each condition ( n = 9). * P < 0.05, relative to the low-K + condition, by 1-way ANOVA followed by multiple-comparison test. Data are presented as the mean ± SEM. CALNA, calcineurin.
Figure Legend Snippet: ( A ) In vitro protein phosphatase (PP) assay. Representative immunoblots of p-NCC and t-NCC are shown for assays of NCC with PP1A (lanes 1 and 2), PP2A (lanes 5 and 6), and calcineurin A (CALNA) or vehicle (lanes 3, 7, and 11). For these studies, NCC was isolated by IP with an anti-NCC antibody (lanes 1, 2, 3, 5, 6, and 7) and compared with the negative control IgG (lanes 4, 8, and 12). ( B ) PP1A preferentially interacted with NCC in glutathione-agarose affinity chromatography assays with the GST fusion protein of the NCC terminus GST-NCC-(N), but not the negative control GST alone. Shown are representative immunoblot binding assays with recombinant PP1A, PP2A, and CALNA. Input protein phosphatase (lane 1) is shown relative to GST-bound (lane 2) or GST-NCC-(N) (lane 3). ( C ) A greater amount of PP1A co-immunoprecipitated with NCC when the extracellular K + concentration was high. Representative immunoblots show NCC, PP1A, and p-NCC (T58) in anti-FLAG immunoprecipitated samples relative to input from FLAG-tagged NCC-expressing MDCKI cells incubated with 3.5 mM (control), 0.5 mM (low), or 8 mM (high) K + buffers. Low-chloride buffer was used as a positive control to elevate p-NCC (T58) levels. Graphs show semiquantitative assessment of NCC, PP1, and p-NCC (T58) in NCC immunoprecipitated samples (bottom, right) relative to input. Data are from 3 individual experiments with 3 replicates for each condition ( n = 9). * P < 0.05, relative to the low-K + condition, by 1-way ANOVA followed by multiple-comparison test. Data are presented as the mean ± SEM. CALNA, calcineurin.

Techniques Used: In Vitro, Western Blot, Isolation, Negative Control, Affinity Chromatography, Binding Assay, Recombinant, Immunoprecipitation, Concentration Assay, Expressing, Incubation, Positive Control, Comparison


Structured Review

Millipore pre washed anti flag m2 magnetic beads
Pre Washed Anti Flag M2 Magnetic Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pre washed anti flag m2 magnetic beads/product/Millipore
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
pre washed anti flag m2 magnetic beads - by Bioz Stars, 2023-11
86/100 stars

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Structured Review

Millipore anti flag m2 affinity gel
Anti Flag M2 Affinity Gel, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti flag m2 affinity gel/product/Millipore
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti flag m2 affinity gel - by Bioz Stars, 2023-11
86/100 stars

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