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  • 97
    Millipore monoclonal anti flag m2 antibody
    CAF-1 subunits interact with <t>Cse4.</t> Whole cell extracts were used to carry out co-IPs in ( A – C ). (A) Left panel: strains carrying GFP-tagged chaperones, Scm3, Vps75, Nap1 and Cac2 subunit of CAF-1 were used in co-IP. Only Cac2-GFP pulled down Cse4. A control IP was performed from a strain lacking the GFP tag. Scm3-GFP IP was the positive control. Right panel: affinity-tagged strains <t>(Cse4-Myc/Cac2-FLAG</t> and Cse4-Myc/Hir1-FLAG) were used for co-IP. A control IP was performed from a strain lacking the FLAG tag. In anti-FLAG co-IP, Hir1-FLAG did not pull down Cse4, only Cac2-FLAG pulled down Cse4-Myc. (B) Cac1 and Cac3 subunits of CAF-1 are required for interaction with Cse4. Upper panel: WT or cac2Δ strains carrying FLAG-tagged Cac1 or Cac3 were used for co-IP. A strain lacking the tag on Cac1 and Cac3 was used as the negative control. In anti-FLAG co-IP, both Cac1 and Cac3 pulled down noticeably more Cse4-Myc. These interactions became weaker with CAC2 deletion. Lower panel: WT, cac1Δ or cac3Δ strains carrying FLAG-tagged Cac2 were used in anti-FLAG co-IP. A strain lacking the tag was used as the control. Cac2/Cse4 interaction was completely abolished in cac1Δ or cac3Δ strains. (C) CAF-1 can interact with Cse4 outside of S phase. Affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Scm3-FLAG) were used in anti-FLAG co-IP using asynchronously grown cells and G2/M-arrested cells. A strain lacking the FLAG tag served as a negative control. Scm3 was used as a positive control. Cac2 interacted with Cse4 in both asynchronously grown cells as well as G2/M-arrested cells. A non-specific band is marked with an asterisk.
    Monoclonal Anti Flag M2 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti flag m2 antibody/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti flag m2 antibody - by Bioz Stars, 2021-03
    97/100 stars
      Buy from Supplier

    99
    Millipore anti flag m2 affinity gel
    CAF-1 subunits interact with <t>Cse4.</t> Whole cell extracts were used to carry out co-IPs in ( A – C ). (A) Left panel: strains carrying GFP-tagged chaperones, Scm3, Vps75, Nap1 and Cac2 subunit of CAF-1 were used in co-IP. Only Cac2-GFP pulled down Cse4. A control IP was performed from a strain lacking the GFP tag. Scm3-GFP IP was the positive control. Right panel: affinity-tagged strains <t>(Cse4-Myc/Cac2-FLAG</t> and Cse4-Myc/Hir1-FLAG) were used for co-IP. A control IP was performed from a strain lacking the FLAG tag. In anti-FLAG co-IP, Hir1-FLAG did not pull down Cse4, only Cac2-FLAG pulled down Cse4-Myc. (B) Cac1 and Cac3 subunits of CAF-1 are required for interaction with Cse4. Upper panel: WT or cac2Δ strains carrying FLAG-tagged Cac1 or Cac3 were used for co-IP. A strain lacking the tag on Cac1 and Cac3 was used as the negative control. In anti-FLAG co-IP, both Cac1 and Cac3 pulled down noticeably more Cse4-Myc. These interactions became weaker with CAC2 deletion. Lower panel: WT, cac1Δ or cac3Δ strains carrying FLAG-tagged Cac2 were used in anti-FLAG co-IP. A strain lacking the tag was used as the control. Cac2/Cse4 interaction was completely abolished in cac1Δ or cac3Δ strains. (C) CAF-1 can interact with Cse4 outside of S phase. Affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Scm3-FLAG) were used in anti-FLAG co-IP using asynchronously grown cells and G2/M-arrested cells. A strain lacking the FLAG tag served as a negative control. Scm3 was used as a positive control. Cac2 interacted with Cse4 in both asynchronously grown cells as well as G2/M-arrested cells. A non-specific band is marked with an asterisk.
    Anti Flag M2 Affinity Gel, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, 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 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti flag m2 affinity gel - by Bioz Stars, 2021-03
    99/100 stars
      Buy from Supplier

    99
    Millipore anti flag antibody
    CAF-1 subunits interact with <t>Cse4.</t> Whole cell extracts were used to carry out co-IPs in ( A – C ). (A) Left panel: strains carrying GFP-tagged chaperones, Scm3, Vps75, Nap1 and Cac2 subunit of CAF-1 were used in co-IP. Only Cac2-GFP pulled down Cse4. A control IP was performed from a strain lacking the GFP tag. Scm3-GFP IP was the positive control. Right panel: affinity-tagged strains <t>(Cse4-Myc/Cac2-FLAG</t> and Cse4-Myc/Hir1-FLAG) were used for co-IP. A control IP was performed from a strain lacking the FLAG tag. In anti-FLAG co-IP, Hir1-FLAG did not pull down Cse4, only Cac2-FLAG pulled down Cse4-Myc. (B) Cac1 and Cac3 subunits of CAF-1 are required for interaction with Cse4. Upper panel: WT or cac2Δ strains carrying FLAG-tagged Cac1 or Cac3 were used for co-IP. A strain lacking the tag on Cac1 and Cac3 was used as the negative control. In anti-FLAG co-IP, both Cac1 and Cac3 pulled down noticeably more Cse4-Myc. These interactions became weaker with CAC2 deletion. Lower panel: WT, cac1Δ or cac3Δ strains carrying FLAG-tagged Cac2 were used in anti-FLAG co-IP. A strain lacking the tag was used as the control. Cac2/Cse4 interaction was completely abolished in cac1Δ or cac3Δ strains. (C) CAF-1 can interact with Cse4 outside of S phase. Affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Scm3-FLAG) were used in anti-FLAG co-IP using asynchronously grown cells and G2/M-arrested cells. A strain lacking the FLAG tag served as a negative control. Scm3 was used as a positive control. Cac2 interacted with Cse4 in both asynchronously grown cells as well as G2/M-arrested cells. A non-specific band is marked with an asterisk.
    Anti Flag Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti flag antibody/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti flag antibody - by Bioz Stars, 2021-03
    99/100 stars
      Buy from Supplier

    N/A
    Mouse monoclonal to Flag Tag Conjugation note Unconjugated Application note WB IP IF
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    N/A
    Mouse monoclonal to Flag KLH Conjugation note KLH Application note WB ELISA
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    N/A
    Eu W1024 labeled anti FLAG antibodies for LANCE TR FRET assays The anti FLAG antibody is a purified mouse IgG2a monoclonal antibody clone 29E4 G7 H11 The antibody is know
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    Image Search Results


    CAF-1 subunits interact with Cse4. Whole cell extracts were used to carry out co-IPs in ( A – C ). (A) Left panel: strains carrying GFP-tagged chaperones, Scm3, Vps75, Nap1 and Cac2 subunit of CAF-1 were used in co-IP. Only Cac2-GFP pulled down Cse4. A control IP was performed from a strain lacking the GFP tag. Scm3-GFP IP was the positive control. Right panel: affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Hir1-FLAG) were used for co-IP. A control IP was performed from a strain lacking the FLAG tag. In anti-FLAG co-IP, Hir1-FLAG did not pull down Cse4, only Cac2-FLAG pulled down Cse4-Myc. (B) Cac1 and Cac3 subunits of CAF-1 are required for interaction with Cse4. Upper panel: WT or cac2Δ strains carrying FLAG-tagged Cac1 or Cac3 were used for co-IP. A strain lacking the tag on Cac1 and Cac3 was used as the negative control. In anti-FLAG co-IP, both Cac1 and Cac3 pulled down noticeably more Cse4-Myc. These interactions became weaker with CAC2 deletion. Lower panel: WT, cac1Δ or cac3Δ strains carrying FLAG-tagged Cac2 were used in anti-FLAG co-IP. A strain lacking the tag was used as the control. Cac2/Cse4 interaction was completely abolished in cac1Δ or cac3Δ strains. (C) CAF-1 can interact with Cse4 outside of S phase. Affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Scm3-FLAG) were used in anti-FLAG co-IP using asynchronously grown cells and G2/M-arrested cells. A strain lacking the FLAG tag served as a negative control. Scm3 was used as a positive control. Cac2 interacted with Cse4 in both asynchronously grown cells as well as G2/M-arrested cells. A non-specific band is marked with an asterisk.

    Journal: Nucleic Acids Research

    Article Title: Chromatin assembly factor-1 (CAF-1) chaperone regulates Cse4 deposition into chromatin in budding yeast

    doi: 10.1093/nar/gky169

    Figure Lengend Snippet: CAF-1 subunits interact with Cse4. Whole cell extracts were used to carry out co-IPs in ( A – C ). (A) Left panel: strains carrying GFP-tagged chaperones, Scm3, Vps75, Nap1 and Cac2 subunit of CAF-1 were used in co-IP. Only Cac2-GFP pulled down Cse4. A control IP was performed from a strain lacking the GFP tag. Scm3-GFP IP was the positive control. Right panel: affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Hir1-FLAG) were used for co-IP. A control IP was performed from a strain lacking the FLAG tag. In anti-FLAG co-IP, Hir1-FLAG did not pull down Cse4, only Cac2-FLAG pulled down Cse4-Myc. (B) Cac1 and Cac3 subunits of CAF-1 are required for interaction with Cse4. Upper panel: WT or cac2Δ strains carrying FLAG-tagged Cac1 or Cac3 were used for co-IP. A strain lacking the tag on Cac1 and Cac3 was used as the negative control. In anti-FLAG co-IP, both Cac1 and Cac3 pulled down noticeably more Cse4-Myc. These interactions became weaker with CAC2 deletion. Lower panel: WT, cac1Δ or cac3Δ strains carrying FLAG-tagged Cac2 were used in anti-FLAG co-IP. A strain lacking the tag was used as the control. Cac2/Cse4 interaction was completely abolished in cac1Δ or cac3Δ strains. (C) CAF-1 can interact with Cse4 outside of S phase. Affinity-tagged strains (Cse4-Myc/Cac2-FLAG and Cse4-Myc/Scm3-FLAG) were used in anti-FLAG co-IP using asynchronously grown cells and G2/M-arrested cells. A strain lacking the FLAG tag served as a negative control. Scm3 was used as a positive control. Cac2 interacted with Cse4 in both asynchronously grown cells as well as G2/M-arrested cells. A non-specific band is marked with an asterisk.

    Article Snippet: Antibodies used for ChIP experiments are 2 μl of α-Cse4 antibody (polyclonal rabbit antibody against recombinant Cse4) and anti-FLAG (Sigma-F3165).

    Techniques: Co-Immunoprecipitation Assay, Positive Control, FLAG-tag, Negative Control

    When Scm3 is absent and Cse4 levels are high, CAF-1 may help target Cse4 to the centromere. ( A ) Cac1 localization to centromeres is cell-cycle independent. ChIP was performed for Cac1 using a FLAG-tagged strain using asynchronously growing cells and also G1, S and G2/M arrested cells. qPCR was used to detect Cac1 levels at CEN3. The y -axis indicates arbitrary units representing the Cac1 enrichment at CEN3 from ChIP performed with and without antibody, with respect to the signal for total chromatin for each sample. The error bars represent the standard deviation for ChIP performed in triplicate. ( B ) Rescue of Scm3 off strain by overexpression of Cse4 is less efficient when CAC2 is deleted. Strains containing a single copy of Scm3 under the control of the gal promoter, allowing the expression of Scm3 to be controlled with glucose/galactose, were used in the spotting assay. Cse4 was under the control of a copper-inducible promoter on a plasmid. ‘EV’ indicates empty vector. When Scm3 is expressed (Scm3 on ), no growth differences are observed in 10-fold serial dilutions. Induction of Cse4 does not efficiently rescue growth of the Scm3 off strain when CAC2 is deleted as compared to WT. Three independent isolates of the cac2Δ strain (1, 2 and 3) were tested.

    Journal: Nucleic Acids Research

    Article Title: Chromatin assembly factor-1 (CAF-1) chaperone regulates Cse4 deposition into chromatin in budding yeast

    doi: 10.1093/nar/gky169

    Figure Lengend Snippet: When Scm3 is absent and Cse4 levels are high, CAF-1 may help target Cse4 to the centromere. ( A ) Cac1 localization to centromeres is cell-cycle independent. ChIP was performed for Cac1 using a FLAG-tagged strain using asynchronously growing cells and also G1, S and G2/M arrested cells. qPCR was used to detect Cac1 levels at CEN3. The y -axis indicates arbitrary units representing the Cac1 enrichment at CEN3 from ChIP performed with and without antibody, with respect to the signal for total chromatin for each sample. The error bars represent the standard deviation for ChIP performed in triplicate. ( B ) Rescue of Scm3 off strain by overexpression of Cse4 is less efficient when CAC2 is deleted. Strains containing a single copy of Scm3 under the control of the gal promoter, allowing the expression of Scm3 to be controlled with glucose/galactose, were used in the spotting assay. Cse4 was under the control of a copper-inducible promoter on a plasmid. ‘EV’ indicates empty vector. When Scm3 is expressed (Scm3 on ), no growth differences are observed in 10-fold serial dilutions. Induction of Cse4 does not efficiently rescue growth of the Scm3 off strain when CAC2 is deleted as compared to WT. Three independent isolates of the cac2Δ strain (1, 2 and 3) were tested.

    Article Snippet: Antibodies used for ChIP experiments are 2 μl of α-Cse4 antibody (polyclonal rabbit antibody against recombinant Cse4) and anti-FLAG (Sigma-F3165).

    Techniques: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Standard Deviation, Over Expression, Expressing, Spotting Assay, Plasmid Preparation