protein g magnetic beads  (New England Biolabs)


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    Name:
    Protein G Magnetic Beads
    Description:
    Protein G Magnetic Beads 1 ml
    Catalog Number:
    s1430s
    Price:
    202
    Size:
    1 ml
    Category:
    Magnetic Separation Equipment
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    New England Biolabs protein g magnetic beads
    Protein G Magnetic Beads
    Protein G Magnetic Beads 1 ml
    https://www.bioz.com/result/protein g magnetic beads/product/New England Biolabs
    Average 94 stars, based on 244 article reviews
    Price from $9.99 to $1999.99
    protein g magnetic beads - by Bioz Stars, 2021-02
    94/100 stars

    Images

    1) Product Images from "Importance of the 1+7 configuration of ribonucleoprotein complexes for influenza A virus genome packaging"

    Article Title: Importance of the 1+7 configuration of ribonucleoprotein complexes for influenza A virus genome packaging

    Journal: Nature Communications

    doi: 10.1038/s41467-017-02517-w

    Co-immunoprecipitation of 18S and 28S rRNA with the NP protein. Purified virions were disrupted in buffer containing TritonX-100, incubated with an anti-NP antibody, and precipitated with Protein G beads. a The precipitates were subjected to western blot analysis using an anti-whole virion polyclonal antibody. b – d RNAs were extracted from the precipitate and were subjected to northern blotting by using riboprobes specific for NS vRNA ( b ), 18S rRNA ( c ), and 28S rRNA ( d ). IP immunoprecipitation
    Figure Legend Snippet: Co-immunoprecipitation of 18S and 28S rRNA with the NP protein. Purified virions were disrupted in buffer containing TritonX-100, incubated with an anti-NP antibody, and precipitated with Protein G beads. a The precipitates were subjected to western blot analysis using an anti-whole virion polyclonal antibody. b – d RNAs were extracted from the precipitate and were subjected to northern blotting by using riboprobes specific for NS vRNA ( b ), 18S rRNA ( c ), and 28S rRNA ( d ). IP immunoprecipitation

    Techniques Used: Immunoprecipitation, Purification, Incubation, Western Blot, Northern Blot

    2) Product Images from "Identification of distinctive interdomain interactions among ZP-N, ZP-C and other domains of zona pellucida glycoproteins underlying association of chicken egg-coat matrix"

    Article Title: Identification of distinctive interdomain interactions among ZP-N, ZP-C and other domains of zona pellucida glycoproteins underlying association of chicken egg-coat matrix

    Journal: FEBS Open Bio

    doi: 10.1016/j.fob.2015.05.005

    Co-immunoprecipitation assays of ZP1 in the chicken serum and ZP3 isolated from the egg coat. The chicken serum containing ZP1 and the ZP3 solution prepared as described in Section 4 were mixed and incubated to form the ZP1–ZP3 complexes. Protein G magnetic beads coated with the antibodies in the anti-ZP1_N-terminal, repeat, trefoil, ZP-N and ZP-C antisera (A) and the anti-ZP3_N-terminal, ZP-N and ZP-C antisera (B) were incubated in the presence (+; odd-numbered lanes, except for lane 11) or absence (−; even-numbered lanes) of the serum–ZP3 mixture and pulled down by magnetic field. The immunoprecipitates on the beads were subjected to SDS–PAGE under reducing conditions, together with the serum and the ZP3 solution (upper and lower panels in lane 11 of A, respectively) followed by immunoblotting with the anti-ZP1_repeat (anti-ZP1) and anti-ZP3_ZP-C (anti-ZP3) antisera (upper and lower panels, respectively). Migration positions of ZP1 (filled arrowheads), ZP3 (blank arrowheads), and immunoglobulin heavy and light chains (single and double asterisks, respectively) are shown on the right. Molecular weights (kDa) are indicated on the left.
    Figure Legend Snippet: Co-immunoprecipitation assays of ZP1 in the chicken serum and ZP3 isolated from the egg coat. The chicken serum containing ZP1 and the ZP3 solution prepared as described in Section 4 were mixed and incubated to form the ZP1–ZP3 complexes. Protein G magnetic beads coated with the antibodies in the anti-ZP1_N-terminal, repeat, trefoil, ZP-N and ZP-C antisera (A) and the anti-ZP3_N-terminal, ZP-N and ZP-C antisera (B) were incubated in the presence (+; odd-numbered lanes, except for lane 11) or absence (−; even-numbered lanes) of the serum–ZP3 mixture and pulled down by magnetic field. The immunoprecipitates on the beads were subjected to SDS–PAGE under reducing conditions, together with the serum and the ZP3 solution (upper and lower panels in lane 11 of A, respectively) followed by immunoblotting with the anti-ZP1_repeat (anti-ZP1) and anti-ZP3_ZP-C (anti-ZP3) antisera (upper and lower panels, respectively). Migration positions of ZP1 (filled arrowheads), ZP3 (blank arrowheads), and immunoglobulin heavy and light chains (single and double asterisks, respectively) are shown on the right. Molecular weights (kDa) are indicated on the left.

    Techniques Used: Immunoprecipitation, Isolation, Incubation, Magnetic Beads, SDS Page, Migration

    3) Product Images from "Botch (NPG7) Promotes Neurogenesis by Antagonizing Notch"

    Article Title: Botch (NPG7) Promotes Neurogenesis by Antagonizing Notch

    Journal: Developmental Cell

    doi: 10.1016/j.devcel.2012.02.011

    Botch preferentially interacts with the Notch1 extracellular domain (A) Immunoblot of co-immunoprecipitation of endogenous uncleaved immature full-length Notch1 by an anti-Botch polyclonal antibody (Botch-PoAb) from E14.5 ganglionic eminence. An unrelated rabbit IgG antibody and preabsorption of the Botch-PoAb with recombinant Botch-GST are negative controls. Abbreviations: IP, immunoprecipitation; FL, Full Length; TMIC, TransMembrane and IntraCellular domain. (B) Immunoblot of co-immunoprecipitation with Botch-PoAb and Dll1, EGFR or FGRF2 from E14.5 ganglionic eminence. (C and D) Immunoblots of Botch co-immunoprecipitation with the Notch ExtraCellular Domain (NECD1) or the Notch IntraCellular Domain (NICD1). (E) Immunoblot for GFP of Botch co-immunoprecipitation with four fragments of the NECD1; EGF repeats 1-12, 11-24, 22-33 and 32-36 with Lin12/Notch repeats (LNR) that encode rat Notch1 amino acids 1-493, 482-910, 832-1310 and 1224-1723 with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (F) Immunoblot for GFP of Botch co-immunoprecipitation with fragments containing EGF repeats 32-36 and LNR divided into EGF repeats 32-36 (rat Notch1 amino acids 1224-1448) or the Lin12/Notch repeats (LNR) (rat Notch1 amino acids 1449-1723) with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (G) Co-immunoprecipitation of Botch with Notch1 (Flag-N1-Gal4) or Notch Loop Out (Flag-N1-Gal4-LO) which lacks the S1-cleavage sites, visualized by immunoblot. Flag-N1-Gal4-LO lacks of amino acid sequence 1624-1670 in human Notch1. (H) Scatchard plot of quantitative binding of Botch-N-AP to Notch1 immobilized by anti-Notch1 on protein G sepharose beads. A-H, Experiments were repeated three times with similar results.
    Figure Legend Snippet: Botch preferentially interacts with the Notch1 extracellular domain (A) Immunoblot of co-immunoprecipitation of endogenous uncleaved immature full-length Notch1 by an anti-Botch polyclonal antibody (Botch-PoAb) from E14.5 ganglionic eminence. An unrelated rabbit IgG antibody and preabsorption of the Botch-PoAb with recombinant Botch-GST are negative controls. Abbreviations: IP, immunoprecipitation; FL, Full Length; TMIC, TransMembrane and IntraCellular domain. (B) Immunoblot of co-immunoprecipitation with Botch-PoAb and Dll1, EGFR or FGRF2 from E14.5 ganglionic eminence. (C and D) Immunoblots of Botch co-immunoprecipitation with the Notch ExtraCellular Domain (NECD1) or the Notch IntraCellular Domain (NICD1). (E) Immunoblot for GFP of Botch co-immunoprecipitation with four fragments of the NECD1; EGF repeats 1-12, 11-24, 22-33 and 32-36 with Lin12/Notch repeats (LNR) that encode rat Notch1 amino acids 1-493, 482-910, 832-1310 and 1224-1723 with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (F) Immunoblot for GFP of Botch co-immunoprecipitation with fragments containing EGF repeats 32-36 and LNR divided into EGF repeats 32-36 (rat Notch1 amino acids 1224-1448) or the Lin12/Notch repeats (LNR) (rat Notch1 amino acids 1449-1723) with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (G) Co-immunoprecipitation of Botch with Notch1 (Flag-N1-Gal4) or Notch Loop Out (Flag-N1-Gal4-LO) which lacks the S1-cleavage sites, visualized by immunoblot. Flag-N1-Gal4-LO lacks of amino acid sequence 1624-1670 in human Notch1. (H) Scatchard plot of quantitative binding of Botch-N-AP to Notch1 immobilized by anti-Notch1 on protein G sepharose beads. A-H, Experiments were repeated three times with similar results.

    Techniques Used: Immunoprecipitation, Recombinant, Western Blot, Sequencing, Binding Assay

    4) Product Images from "Modification of Adenosine196 by Mettl3 Methyltransferase in the 5’-External Transcribed Spacer of 47S Pre-rRNA Affects rRNA Maturation"

    Article Title: Modification of Adenosine196 by Mettl3 Methyltransferase in the 5’-External Transcribed Spacer of 47S Pre-rRNA Affects rRNA Maturation

    Journal: Cells

    doi: 10.3390/cells9041061

    Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. ( A ) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. ( B ) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p
    Figure Legend Snippet: Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. ( A ) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. ( B ) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p

    Techniques Used: Quantitative RT-PCR

    5) Product Images from "Glycosylation profiling of dog serum reveals differences compared to human serum"

    Article Title: Glycosylation profiling of dog serum reveals differences compared to human serum

    Journal: Glycobiology

    doi: 10.1093/glycob/cwy070

    ( A ) HILIC-UPLC profiles of enzymatically released and procainamide-labeled N -glycans from human (blue, top) and canine (green, bottom) blood serum. ( B ) HILIC-UPLC profile of N -glycans released from canine IgG. IgG was purified using Protein G from canine serum. Glycan structures are annotated following the nomenclature outlined by the Consortium for Functional Glycomics (CFG). The inset in B shows the monosaccharide symbols.
    Figure Legend Snippet: ( A ) HILIC-UPLC profiles of enzymatically released and procainamide-labeled N -glycans from human (blue, top) and canine (green, bottom) blood serum. ( B ) HILIC-UPLC profile of N -glycans released from canine IgG. IgG was purified using Protein G from canine serum. Glycan structures are annotated following the nomenclature outlined by the Consortium for Functional Glycomics (CFG). The inset in B shows the monosaccharide symbols.

    Techniques Used: Hydrophilic Interaction Liquid Chromatography, Labeling, Purification, Functional Assay

    6) Product Images from "HIV envelope-mediated, CCR5/?4?7-dependent killing of CD4-negative ?? T cells which are lost during progression to AIDS"

    Article Title: HIV envelope-mediated, CCR5/?4?7-dependent killing of CD4-negative ?? T cells which are lost during progression to AIDS

    Journal: Blood

    doi: 10.1182/blood-2011-05-356535

    α4β7 and CCR5 form complexes on Vγ2Vδ2 T cells. (A) Flow cytometry assay for expression of CCR5 and β7 on Vγ2Vδ2 T cells. (B) Vγ2Vδ2 T cells were stained with β7 mAb (green) and CCR5 mAb (red), and viewed under a confocal microscope. Yellow in the merged panel represents the colocalization of CCR5 and β7. (C) Vγ2Vδ2 T cells were treated or not with the crosslinking reagent DTSSP followed by coprecipitation with protein G magnetic beads and the rat IgG2a+DTSSP (lane 1), β7 mAb FIB27 (lane 2), β7 mAb FIB27 + DTSSP (lane 3). Cell lysates were run as a positive control (lane 4). CCR5 and β7 were detected by Western blot with specific antibodies. Data are representative of 3 independent experiments.
    Figure Legend Snippet: α4β7 and CCR5 form complexes on Vγ2Vδ2 T cells. (A) Flow cytometry assay for expression of CCR5 and β7 on Vγ2Vδ2 T cells. (B) Vγ2Vδ2 T cells were stained with β7 mAb (green) and CCR5 mAb (red), and viewed under a confocal microscope. Yellow in the merged panel represents the colocalization of CCR5 and β7. (C) Vγ2Vδ2 T cells were treated or not with the crosslinking reagent DTSSP followed by coprecipitation with protein G magnetic beads and the rat IgG2a+DTSSP (lane 1), β7 mAb FIB27 (lane 2), β7 mAb FIB27 + DTSSP (lane 3). Cell lysates were run as a positive control (lane 4). CCR5 and β7 were detected by Western blot with specific antibodies. Data are representative of 3 independent experiments.

    Techniques Used: Flow Cytometry, Cytometry, Expressing, Staining, Microscopy, Magnetic Beads, Positive Control, Western Blot

    7) Product Images from "Characterization of the interaction between Toxoplasma gondii rhoptry neck protein 4 and host cellular ?-tubulin"

    Article Title: Characterization of the interaction between Toxoplasma gondii rhoptry neck protein 4 and host cellular ?-tubulin

    Journal: Scientific Reports

    doi: 10.1038/srep03199

    Identification of TgRON4-binding host proteins. (a) Recombinant proteins expressed using the baculovirus expression system and purified on Ni-NTA agarose. Each protein (50 ng) was separated by 5%–20% gradient SDS-PAGE and subjected to silver staining (lanes 2–4). The molecular masses (kDa) are indicated on the left. Immunoblotting with an HRP-conjugated anti-mouse Fc antibody of purified recombinant proteins (lanes 5–7). (b) Each Fc-fusion protein was crosslinked to protein G magnetic beads and incubated with membrane proteins from 293 T cells. The eluate was separated by SDS-PAGE followed by silver staining (lanes 1–3). The arrowhead indicates a 50-kDa band that is specific for incubation with TgRON4-linked beads. Immunoblotting of the eluates with an anti-β-tubulin antibody (lanes 4–6).
    Figure Legend Snippet: Identification of TgRON4-binding host proteins. (a) Recombinant proteins expressed using the baculovirus expression system and purified on Ni-NTA agarose. Each protein (50 ng) was separated by 5%–20% gradient SDS-PAGE and subjected to silver staining (lanes 2–4). The molecular masses (kDa) are indicated on the left. Immunoblotting with an HRP-conjugated anti-mouse Fc antibody of purified recombinant proteins (lanes 5–7). (b) Each Fc-fusion protein was crosslinked to protein G magnetic beads and incubated with membrane proteins from 293 T cells. The eluate was separated by SDS-PAGE followed by silver staining (lanes 1–3). The arrowhead indicates a 50-kDa band that is specific for incubation with TgRON4-linked beads. Immunoblotting of the eluates with an anti-β-tubulin antibody (lanes 4–6).

    Techniques Used: Binding Assay, Recombinant, Expressing, Purification, SDS Page, Silver Staining, Magnetic Beads, Incubation

    8) Product Images from "Inhibition of Wnt/?-Catenin Signaling by a Soluble Collagen-Derived Frizzled Domain Interacting with Wnt3a and the Receptors Frizzled 1 and 8"

    Article Title: Inhibition of Wnt/?-Catenin Signaling by a Soluble Collagen-Derived Frizzled Domain Interacting with Wnt3a and the Receptors Frizzled 1 and 8

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030601

    The FZC18 domain homodimerizes and binds FZD1 and FZD8 CRDs. (A) Schematic structure of V3Nter, V2Nter and FZC18 cDNAs. V3Nter and V2Nter correspond to the N-terminal noncollagenous domains of variants 3 and 2 of collagen XVIII, respectively. They share the DUF-959 domain, a portion of the tsp-1 (thrombospondin-1) domain and the V5 tag. Only V3Nter contains the FZC18 domain. The FZC18 vector has a myc tag. Thick horizontal lines indicate the antibodies used. (B) FZC18 can homodimerize. FZC18-myc was cotransfected with V3Nter-V5 or V2Nter-V5 in HEK293-EBNA cells. Cell lysates were immunoprecipitated with anti-myc and immunoblotted with anti-DUF-959 (top) . The membrane was stripped and re-probed with anti-V5 (bottom) . Ig , immunoglobulins. (C) Soluble FZC18 binds FZD1_CRD and FZD8_CRD. CM from HEK293-EBNA cells secreting FZC18-myc was incubated with recombinant 100 ng/ml FZD1_CRD-Fc (upper panel) or with CM from HEK293-EBNA cells secreting FZD8_CRD-Fc (lower panel) . FZD1_CRD-Fc and FZD8_CRD-Fc were immunoprecipitated with protein G magnetic beads, electrophoresed and immunoblotted with anti-myc, anti-FZD1_CRD or anti-FZD8_CRD, as shown. Asterisks denote inputs or FZC18 cell lysate, as indicated.
    Figure Legend Snippet: The FZC18 domain homodimerizes and binds FZD1 and FZD8 CRDs. (A) Schematic structure of V3Nter, V2Nter and FZC18 cDNAs. V3Nter and V2Nter correspond to the N-terminal noncollagenous domains of variants 3 and 2 of collagen XVIII, respectively. They share the DUF-959 domain, a portion of the tsp-1 (thrombospondin-1) domain and the V5 tag. Only V3Nter contains the FZC18 domain. The FZC18 vector has a myc tag. Thick horizontal lines indicate the antibodies used. (B) FZC18 can homodimerize. FZC18-myc was cotransfected with V3Nter-V5 or V2Nter-V5 in HEK293-EBNA cells. Cell lysates were immunoprecipitated with anti-myc and immunoblotted with anti-DUF-959 (top) . The membrane was stripped and re-probed with anti-V5 (bottom) . Ig , immunoglobulins. (C) Soluble FZC18 binds FZD1_CRD and FZD8_CRD. CM from HEK293-EBNA cells secreting FZC18-myc was incubated with recombinant 100 ng/ml FZD1_CRD-Fc (upper panel) or with CM from HEK293-EBNA cells secreting FZD8_CRD-Fc (lower panel) . FZD1_CRD-Fc and FZD8_CRD-Fc were immunoprecipitated with protein G magnetic beads, electrophoresed and immunoblotted with anti-myc, anti-FZD1_CRD or anti-FZD8_CRD, as shown. Asterisks denote inputs or FZC18 cell lysate, as indicated.

    Techniques Used: Plasmid Preparation, Immunoprecipitation, Incubation, Recombinant, Magnetic Beads

    9) Product Images from "Estradiol-Estrogen Receptor α Mediates the Expression of the CXXC5 Gene through the Estrogen Response Element-Dependent Signaling Pathway"

    Article Title: Estradiol-Estrogen Receptor α Mediates the Expression of the CXXC5 Gene through the Estrogen Response Element-Dependent Signaling Pathway

    Journal: Scientific Reports

    doi: 10.1038/srep37808

    Chromatin Immunoprecipitation assay (ChIP). MCF7 cells grown in medium containing CD-FBS for 72 h treated without (EtOH, 0.01%) with 10 −8  M E2 for 1 h prior to ChIP. Cells were fixed with 0.75% paraformaldehyde, lysed, sonicated and subjected to ChIP using IgG or an ERα specific HC20x antibody followed by the incubation with Protein A/G conjugated magnetic beads. Shown ( a ) are PCR reactions subjected to 2% agarose gel electrophoresis from a representative experiment performed three independent times. ( b ) Samples were also subjected to RT-qPCR for quantitative analysis with primers specific to the estrogen responsive region of  CXXC5 . ( c ) RT-qPCR results the estrogen responsive region of  TFF1  with the same experimental inputs described in  (b)  with primers specific to the estrogen responsive region of  TFF1 . Sizes of the DNA fragments in base pairs are indicated. Asterisk (*) denotes significant change depicted as percent (%) of input.
    Figure Legend Snippet: Chromatin Immunoprecipitation assay (ChIP). MCF7 cells grown in medium containing CD-FBS for 72 h treated without (EtOH, 0.01%) with 10 −8  M E2 for 1 h prior to ChIP. Cells were fixed with 0.75% paraformaldehyde, lysed, sonicated and subjected to ChIP using IgG or an ERα specific HC20x antibody followed by the incubation with Protein A/G conjugated magnetic beads. Shown ( a ) are PCR reactions subjected to 2% agarose gel electrophoresis from a representative experiment performed three independent times. ( b ) Samples were also subjected to RT-qPCR for quantitative analysis with primers specific to the estrogen responsive region of CXXC5 . ( c ) RT-qPCR results the estrogen responsive region of TFF1 with the same experimental inputs described in (b) with primers specific to the estrogen responsive region of TFF1 . Sizes of the DNA fragments in base pairs are indicated. Asterisk (*) denotes significant change depicted as percent (%) of input.

    Techniques Used: Chromatin Immunoprecipitation, Sonication, Incubation, Magnetic Beads, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Quantitative RT-PCR

    10) Product Images from "Combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes (COMPARE-MS) for the rapid, sensitive and quantitative detection of DNA methylation"

    Article Title: Combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes (COMPARE-MS) for the rapid, sensitive and quantitative detection of DNA methylation

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gnj022

    COMPARE-MS assay performance. ( A and B ) Plots showing the measured amount of methylated GSTP1 CGIs in M.SssI treated and untreated WBC genomic DNA versus the amount of input DNA after enriching for methylated DNA by methylation-sensitive restriction enzymes alone (A) or MBD2-MBD capture alone (B). ( C ) Plot of relative enrichment of M.SssI treated or untreated WBC DNA with or without MBD2-MBD, anti-His antibody and protein G magnetic beads. The degree of capture of unmethylated DNA (untreated WBC DNA) in the presence of MBD2-MBD is less than or equal to the capture of DNA in the absence of MBD2-MBD or anti-His antibody, showing that capture of unmethylated DNA during the DNA capture step of COMPARE-MS is almost completely due to low amounts of non-specific binding to the protein G magnetic beads, as opposed to low-level binding of the MBD2-MBD to unmethylated DNA. ( D ) Plots showing the measured amount of methylated GSTP1 CGIs in M.SssI treated and untreated WBC genomic DNA versus the amount of input DNA after enriching for methylated DNA by the combination of methylation-sensitive restriction enzyme digestion and MBD2-MBD capture (COMPARE-MS) followed by real-time PCR. When 20–100 ng of input DNA are used, COMPARE-MS has a > 5000-fold dynamic range, which is ∼500-fold higher than that of methylation-sensitive restriction enzyme used alone and ∼5- to 10-fold higher than that of MBD2-MBD capture used alone. ( E ) Plot showing measured output methylated GSTP1 CGIs as determined by COMPARE-MS when decreasing amounts of M.SssI-treated WBC DNA is diluted in 20 ng of untreated WBC genomic DNA. The dashed line is a reference representing the mean COMPARE-MS output (0.0065 ± 0.0023 ng) when four identical replicates of 100 ng of untreated, unmixed WBC genomic DNA were analyzed. COMPARE-MS performance in this series of simulated heterogeneous samples (E) is highly linear for almost four orders of magnitude and nearly identical to that seen with homogeneously methylated samples (D), showing robust reproducibility and sensitivity. Data in (A–E) represent mean ± SEM for triplicate measurements.
    Figure Legend Snippet: COMPARE-MS assay performance. ( A and B ) Plots showing the measured amount of methylated GSTP1 CGIs in M.SssI treated and untreated WBC genomic DNA versus the amount of input DNA after enriching for methylated DNA by methylation-sensitive restriction enzymes alone (A) or MBD2-MBD capture alone (B). ( C ) Plot of relative enrichment of M.SssI treated or untreated WBC DNA with or without MBD2-MBD, anti-His antibody and protein G magnetic beads. The degree of capture of unmethylated DNA (untreated WBC DNA) in the presence of MBD2-MBD is less than or equal to the capture of DNA in the absence of MBD2-MBD or anti-His antibody, showing that capture of unmethylated DNA during the DNA capture step of COMPARE-MS is almost completely due to low amounts of non-specific binding to the protein G magnetic beads, as opposed to low-level binding of the MBD2-MBD to unmethylated DNA. ( D ) Plots showing the measured amount of methylated GSTP1 CGIs in M.SssI treated and untreated WBC genomic DNA versus the amount of input DNA after enriching for methylated DNA by the combination of methylation-sensitive restriction enzyme digestion and MBD2-MBD capture (COMPARE-MS) followed by real-time PCR. When 20–100 ng of input DNA are used, COMPARE-MS has a > 5000-fold dynamic range, which is ∼500-fold higher than that of methylation-sensitive restriction enzyme used alone and ∼5- to 10-fold higher than that of MBD2-MBD capture used alone. ( E ) Plot showing measured output methylated GSTP1 CGIs as determined by COMPARE-MS when decreasing amounts of M.SssI-treated WBC DNA is diluted in 20 ng of untreated WBC genomic DNA. The dashed line is a reference representing the mean COMPARE-MS output (0.0065 ± 0.0023 ng) when four identical replicates of 100 ng of untreated, unmixed WBC genomic DNA were analyzed. COMPARE-MS performance in this series of simulated heterogeneous samples (E) is highly linear for almost four orders of magnitude and nearly identical to that seen with homogeneously methylated samples (D), showing robust reproducibility and sensitivity. Data in (A–E) represent mean ± SEM for triplicate measurements.

    Techniques Used: Mass Spectrometry, Methylation, DNA Methylation Assay, Magnetic Beads, Binding Assay, Real-time Polymerase Chain Reaction

    11) Product Images from "Modification of Adenosine196 by Mettl3 Methyltransferase in the 5’-External Transcribed Spacer of 47S Pre-rRNA Affects rRNA Maturation"

    Article Title: Modification of Adenosine196 by Mettl3 Methyltransferase in the 5’-External Transcribed Spacer of 47S Pre-rRNA Affects rRNA Maturation

    Journal: Cells

    doi: 10.3390/cells9041061

    Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. ( A ) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. ( B ) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p
    Figure Legend Snippet: Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. ( A ) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. ( B ) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p

    Techniques Used: Quantitative RT-PCR

    12) Product Images from "Soluble CD36 Ectodomain Binds Negatively Charged Diacylglycerol Ligands and Acts as a Co-Receptor for TLR2"

    Article Title: Soluble CD36 Ectodomain Binds Negatively Charged Diacylglycerol Ligands and Acts as a Co-Receptor for TLR2

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007411

    mCD36ED and TLR2. (A) mCD36ED does not interact with an mTLR2ED/Fc chimera. SDS PAGE of the samples from immunoprecipitation using protein G beads. Lanes: 1- Protein G incubated with mTLR2ED/Fc chimera and mCD36ED; 2- Protein G incubated with mTLR2ED/Fc chimera alone; 3- Protein G incubated with mCD36ED; 4- mCD36ED; 5- mTLR2ED. (B) Model of CD36-dependent activation of the TLR2 signaling pathway. CD36 binds LTA (step 1) which is transferred to CD14 (step 2). Alternatively, the soluble ectodomain of CD36 binds LTA (step 1a), and transfers it to CD14 (step 2a). Subsequently, CD14 transfers LTA to TLR2/TLR6 (step 3) and the MyD88 pathway is activated (step 4).
    Figure Legend Snippet: mCD36ED and TLR2. (A) mCD36ED does not interact with an mTLR2ED/Fc chimera. SDS PAGE of the samples from immunoprecipitation using protein G beads. Lanes: 1- Protein G incubated with mTLR2ED/Fc chimera and mCD36ED; 2- Protein G incubated with mTLR2ED/Fc chimera alone; 3- Protein G incubated with mCD36ED; 4- mCD36ED; 5- mTLR2ED. (B) Model of CD36-dependent activation of the TLR2 signaling pathway. CD36 binds LTA (step 1) which is transferred to CD14 (step 2). Alternatively, the soluble ectodomain of CD36 binds LTA (step 1a), and transfers it to CD14 (step 2a). Subsequently, CD14 transfers LTA to TLR2/TLR6 (step 3) and the MyD88 pathway is activated (step 4).

    Techniques Used: SDS Page, Immunoprecipitation, Incubation, Activation Assay

    13) Product Images from "Mucosal fluid glycoprotein DMBT1 suppresses twitching motility and virulence of the opportunistic pathogen Pseudomonas aeruginosa"

    Article Title: Mucosal fluid glycoprotein DMBT1 suppresses twitching motility and virulence of the opportunistic pathogen Pseudomonas aeruginosa

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006392

    Identification of DMBT1 as the tear fluid inhibitor of P . aeruginosa twitching motility. (A) Human tear fluid was separated into 7 fractions using size exclusion chromatography. (B) Effect of tear fractions on twitching velocity of PAO1 reveals a high Mw fraction retains inhibitory activity. (C) Mass spectrometric analysis of high Mw tear fractions from two size-exclusion experiments reveal 4 proteins common to fractions inhibiting twitching motility. (D) DMBT1-depleted human tear fluid does not inhibit twitching motility of PAO1. (E) Western blot analysis of samples used in (D) shows depletion of DMBT-1 from tear fluid, and partial depletion by isotype control and protein G only beads control. Data shown in panels B and D as mean ± SEM from three independent experiments. Significance was determined using a one-way ANOVA with Tukey's post-hoc analysis. ****, P
    Figure Legend Snippet: Identification of DMBT1 as the tear fluid inhibitor of P . aeruginosa twitching motility. (A) Human tear fluid was separated into 7 fractions using size exclusion chromatography. (B) Effect of tear fractions on twitching velocity of PAO1 reveals a high Mw fraction retains inhibitory activity. (C) Mass spectrometric analysis of high Mw tear fractions from two size-exclusion experiments reveal 4 proteins common to fractions inhibiting twitching motility. (D) DMBT1-depleted human tear fluid does not inhibit twitching motility of PAO1. (E) Western blot analysis of samples used in (D) shows depletion of DMBT-1 from tear fluid, and partial depletion by isotype control and protein G only beads control. Data shown in panels B and D as mean ± SEM from three independent experiments. Significance was determined using a one-way ANOVA with Tukey's post-hoc analysis. ****, P

    Techniques Used: Size-exclusion Chromatography, Activity Assay, Western Blot

    14) Product Images from "Modification of Adenosine196 by Mettl3 Methyltransferase in the 5’-External Transcribed Spacer of 47S Pre-rRNA Affects rRNA Maturation"

    Article Title: Modification of Adenosine196 by Mettl3 Methyltransferase in the 5’-External Transcribed Spacer of 47S Pre-rRNA Affects rRNA Maturation

    Journal: Cells

    doi: 10.3390/cells9041061

    Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. ( A ) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. ( B ) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p
    Figure Legend Snippet: Characterization of RNA methyltransferase Mettl3 interactions with 5′-ETS of 47S pre-rRNA. ( A ) Scheme of the RT-qPCR primer locations on the pre-47S rRNA. ( B ) Relative amounts of rRNA fragments in immunoprecipitates obtained with anti-m6A antibodies from control and Mettl3 KD cells, quantified by RT-qPCR analysis (* p

    Techniques Used: Quantitative RT-PCR

    15) Product Images from "Glycosylation profiling of dog serum reveals differences compared to human serum"

    Article Title: Glycosylation profiling of dog serum reveals differences compared to human serum

    Journal: Glycobiology

    doi: 10.1093/glycob/cwy070

    ( A ) HILIC-UPLC profiles of enzymatically released and procainamide-labeled N -glycans from human (blue, top) and canine (green, bottom) blood serum. ( B ) HILIC-UPLC profile of N -glycans released from canine IgG. IgG was purified using Protein G from canine serum. Glycan structures are annotated following the nomenclature outlined by the Consortium for Functional Glycomics (CFG). The inset in B shows the monosaccharide symbols.
    Figure Legend Snippet: ( A ) HILIC-UPLC profiles of enzymatically released and procainamide-labeled N -glycans from human (blue, top) and canine (green, bottom) blood serum. ( B ) HILIC-UPLC profile of N -glycans released from canine IgG. IgG was purified using Protein G from canine serum. Glycan structures are annotated following the nomenclature outlined by the Consortium for Functional Glycomics (CFG). The inset in B shows the monosaccharide symbols.

    Techniques Used: Hydrophilic Interaction Liquid Chromatography, Labeling, Purification, Functional Assay

    16) Product Images from "Plasmid replication-associated single-strand-specific methyltransferases"

    Article Title: Plasmid replication-associated single-strand-specific methyltransferases

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkaa1163

    Polymerase and MTase activities copurify when domains are fused. Panel ( A ): Size and purity of fusion proteins. For each MTase, both of the immunoreactive components of the MTase-PolI fusion proteins run at the same position, and comigrate with the Coomassie-stained purified proteins. Western blot (lanes 1, 5, 9 and 10) detected 1 μg of MTase-PolI fusion proteins; Coomassie (lanes 2, 3, 6, 7) visualized 1 μg or 20 μg of the same fractions. Western blots were probed separately with anti-Pol1 rabbit polyclonal or anti-6xHis (detecting the MTase) monoclonal antibodies and developed with horseradish peroxidase-labeled antirabbit or antimouse following kit instructions as detailed in Material and Methods. Dots on lane 1 correspond to the position of protein markers after Western blotting. The bands at the side of lane 10 are spillover from the adjacent lane, which were control 6xHis tagged proteins from a PurExpress extract. Panel ( B ): Activity copurification through two columns. Pooled HiTrapHepHP (#22–26) and HiTrapQHP (#15–19) protein fractions were tested for MTase activity on single-stranded M13mp18 DNA in the presence of [H 3 ]SAM and for DNA-polymerase activity on sonicated sperm-whale DNA in the presence of [H 3 ]TTP.
    Figure Legend Snippet: Polymerase and MTase activities copurify when domains are fused. Panel ( A ): Size and purity of fusion proteins. For each MTase, both of the immunoreactive components of the MTase-PolI fusion proteins run at the same position, and comigrate with the Coomassie-stained purified proteins. Western blot (lanes 1, 5, 9 and 10) detected 1 μg of MTase-PolI fusion proteins; Coomassie (lanes 2, 3, 6, 7) visualized 1 μg or 20 μg of the same fractions. Western blots were probed separately with anti-Pol1 rabbit polyclonal or anti-6xHis (detecting the MTase) monoclonal antibodies and developed with horseradish peroxidase-labeled antirabbit or antimouse following kit instructions as detailed in Material and Methods. Dots on lane 1 correspond to the position of protein markers after Western blotting. The bands at the side of lane 10 are spillover from the adjacent lane, which were control 6xHis tagged proteins from a PurExpress extract. Panel ( B ): Activity copurification through two columns. Pooled HiTrapHepHP (#22–26) and HiTrapQHP (#15–19) protein fractions were tested for MTase activity on single-stranded M13mp18 DNA in the presence of [H 3 ]SAM and for DNA-polymerase activity on sonicated sperm-whale DNA in the presence of [H 3 ]TTP.

    Techniques Used: Staining, Purification, Western Blot, Labeling, Activity Assay, Copurification, Sonication

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    Magnetic Beads:

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    Article Title: Botch (NPG7) Promotes Neurogenesis by Antagonizing Notch
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    Blocking Assay:

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    Incubation:

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    New England Biolabs protein g magnetic beads
    Co-immunoprecipitation of 18S and 28S rRNA with the NP protein. Purified virions were disrupted in buffer containing TritonX-100, incubated with an anti-NP antibody, and precipitated with <t>Protein</t> G beads. a The precipitates were subjected to western blot analysis using an anti-whole virion polyclonal antibody. b – d RNAs were extracted from the precipitate and were subjected to northern blotting by using riboprobes specific for NS vRNA ( b ), 18S rRNA ( c ), and 28S rRNA ( d ). IP immunoprecipitation
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    Co-immunoprecipitation of 18S and 28S rRNA with the NP protein. Purified virions were disrupted in buffer containing TritonX-100, incubated with an anti-NP antibody, and precipitated with Protein G beads. a The precipitates were subjected to western blot analysis using an anti-whole virion polyclonal antibody. b – d RNAs were extracted from the precipitate and were subjected to northern blotting by using riboprobes specific for NS vRNA ( b ), 18S rRNA ( c ), and 28S rRNA ( d ). IP immunoprecipitation

    Journal: Nature Communications

    Article Title: Importance of the 1+7 configuration of ribonucleoprotein complexes for influenza A virus genome packaging

    doi: 10.1038/s41467-017-02517-w

    Figure Lengend Snippet: Co-immunoprecipitation of 18S and 28S rRNA with the NP protein. Purified virions were disrupted in buffer containing TritonX-100, incubated with an anti-NP antibody, and precipitated with Protein G beads. a The precipitates were subjected to western blot analysis using an anti-whole virion polyclonal antibody. b – d RNAs were extracted from the precipitate and were subjected to northern blotting by using riboprobes specific for NS vRNA ( b ), 18S rRNA ( c ), and 28S rRNA ( d ). IP immunoprecipitation

    Article Snippet: The lysate was incubated with an anti-NP (2S347/3) mouse monoclonal antibody and Protein G Magnetic Beads (NEB).

    Techniques: Immunoprecipitation, Purification, Incubation, Western Blot, Northern Blot

    Co-immunoprecipitation assays of ZP1 in the chicken serum and ZP3 isolated from the egg coat. The chicken serum containing ZP1 and the ZP3 solution prepared as described in Section 4 were mixed and incubated to form the ZP1–ZP3 complexes. Protein G magnetic beads coated with the antibodies in the anti-ZP1_N-terminal, repeat, trefoil, ZP-N and ZP-C antisera (A) and the anti-ZP3_N-terminal, ZP-N and ZP-C antisera (B) were incubated in the presence (+; odd-numbered lanes, except for lane 11) or absence (−; even-numbered lanes) of the serum–ZP3 mixture and pulled down by magnetic field. The immunoprecipitates on the beads were subjected to SDS–PAGE under reducing conditions, together with the serum and the ZP3 solution (upper and lower panels in lane 11 of A, respectively) followed by immunoblotting with the anti-ZP1_repeat (anti-ZP1) and anti-ZP3_ZP-C (anti-ZP3) antisera (upper and lower panels, respectively). Migration positions of ZP1 (filled arrowheads), ZP3 (blank arrowheads), and immunoglobulin heavy and light chains (single and double asterisks, respectively) are shown on the right. Molecular weights (kDa) are indicated on the left.

    Journal: FEBS Open Bio

    Article Title: Identification of distinctive interdomain interactions among ZP-N, ZP-C and other domains of zona pellucida glycoproteins underlying association of chicken egg-coat matrix

    doi: 10.1016/j.fob.2015.05.005

    Figure Lengend Snippet: Co-immunoprecipitation assays of ZP1 in the chicken serum and ZP3 isolated from the egg coat. The chicken serum containing ZP1 and the ZP3 solution prepared as described in Section 4 were mixed and incubated to form the ZP1–ZP3 complexes. Protein G magnetic beads coated with the antibodies in the anti-ZP1_N-terminal, repeat, trefoil, ZP-N and ZP-C antisera (A) and the anti-ZP3_N-terminal, ZP-N and ZP-C antisera (B) were incubated in the presence (+; odd-numbered lanes, except for lane 11) or absence (−; even-numbered lanes) of the serum–ZP3 mixture and pulled down by magnetic field. The immunoprecipitates on the beads were subjected to SDS–PAGE under reducing conditions, together with the serum and the ZP3 solution (upper and lower panels in lane 11 of A, respectively) followed by immunoblotting with the anti-ZP1_repeat (anti-ZP1) and anti-ZP3_ZP-C (anti-ZP3) antisera (upper and lower panels, respectively). Migration positions of ZP1 (filled arrowheads), ZP3 (blank arrowheads), and immunoglobulin heavy and light chains (single and double asterisks, respectively) are shown on the right. Molecular weights (kDa) are indicated on the left.

    Article Snippet: 4.7 Co-immunoprecipitation assay Protein G magnetic beads (25 μl of 50% slurry; New England Biolabs, Beverly, MA) were prepared according to the manufacturer’s instructions.

    Techniques: Immunoprecipitation, Isolation, Incubation, Magnetic Beads, SDS Page, Migration

    Botch preferentially interacts with the Notch1 extracellular domain (A) Immunoblot of co-immunoprecipitation of endogenous uncleaved immature full-length Notch1 by an anti-Botch polyclonal antibody (Botch-PoAb) from E14.5 ganglionic eminence. An unrelated rabbit IgG antibody and preabsorption of the Botch-PoAb with recombinant Botch-GST are negative controls. Abbreviations: IP, immunoprecipitation; FL, Full Length; TMIC, TransMembrane and IntraCellular domain. (B) Immunoblot of co-immunoprecipitation with Botch-PoAb and Dll1, EGFR or FGRF2 from E14.5 ganglionic eminence. (C and D) Immunoblots of Botch co-immunoprecipitation with the Notch ExtraCellular Domain (NECD1) or the Notch IntraCellular Domain (NICD1). (E) Immunoblot for GFP of Botch co-immunoprecipitation with four fragments of the NECD1; EGF repeats 1-12, 11-24, 22-33 and 32-36 with Lin12/Notch repeats (LNR) that encode rat Notch1 amino acids 1-493, 482-910, 832-1310 and 1224-1723 with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (F) Immunoblot for GFP of Botch co-immunoprecipitation with fragments containing EGF repeats 32-36 and LNR divided into EGF repeats 32-36 (rat Notch1 amino acids 1224-1448) or the Lin12/Notch repeats (LNR) (rat Notch1 amino acids 1449-1723) with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (G) Co-immunoprecipitation of Botch with Notch1 (Flag-N1-Gal4) or Notch Loop Out (Flag-N1-Gal4-LO) which lacks the S1-cleavage sites, visualized by immunoblot. Flag-N1-Gal4-LO lacks of amino acid sequence 1624-1670 in human Notch1. (H) Scatchard plot of quantitative binding of Botch-N-AP to Notch1 immobilized by anti-Notch1 on protein G sepharose beads. A-H, Experiments were repeated three times with similar results.

    Journal: Developmental Cell

    Article Title: Botch (NPG7) Promotes Neurogenesis by Antagonizing Notch

    doi: 10.1016/j.devcel.2012.02.011

    Figure Lengend Snippet: Botch preferentially interacts with the Notch1 extracellular domain (A) Immunoblot of co-immunoprecipitation of endogenous uncleaved immature full-length Notch1 by an anti-Botch polyclonal antibody (Botch-PoAb) from E14.5 ganglionic eminence. An unrelated rabbit IgG antibody and preabsorption of the Botch-PoAb with recombinant Botch-GST are negative controls. Abbreviations: IP, immunoprecipitation; FL, Full Length; TMIC, TransMembrane and IntraCellular domain. (B) Immunoblot of co-immunoprecipitation with Botch-PoAb and Dll1, EGFR or FGRF2 from E14.5 ganglionic eminence. (C and D) Immunoblots of Botch co-immunoprecipitation with the Notch ExtraCellular Domain (NECD1) or the Notch IntraCellular Domain (NICD1). (E) Immunoblot for GFP of Botch co-immunoprecipitation with four fragments of the NECD1; EGF repeats 1-12, 11-24, 22-33 and 32-36 with Lin12/Notch repeats (LNR) that encode rat Notch1 amino acids 1-493, 482-910, 832-1310 and 1224-1723 with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (F) Immunoblot for GFP of Botch co-immunoprecipitation with fragments containing EGF repeats 32-36 and LNR divided into EGF repeats 32-36 (rat Notch1 amino acids 1224-1448) or the Lin12/Notch repeats (LNR) (rat Notch1 amino acids 1449-1723) with cDNA encoding the 20 amino-acid signal peptide from Notch1 at the N-terminal for proper subcellular localization. (G) Co-immunoprecipitation of Botch with Notch1 (Flag-N1-Gal4) or Notch Loop Out (Flag-N1-Gal4-LO) which lacks the S1-cleavage sites, visualized by immunoblot. Flag-N1-Gal4-LO lacks of amino acid sequence 1624-1670 in human Notch1. (H) Scatchard plot of quantitative binding of Botch-N-AP to Notch1 immobilized by anti-Notch1 on protein G sepharose beads. A-H, Experiments were repeated three times with similar results.

    Article Snippet: The Flag-Notch1-GFP protein binding on protein G beads was first pre-treated with AP or Botch-AP, and then treated with furin (New England Biolabs) and DMSO or furin with 50μM DEC-RVKR-CMK (Enzo LifeSciences).

    Techniques: Immunoprecipitation, Recombinant, Western Blot, Sequencing, Binding Assay

    ( A ) HILIC-UPLC profiles of enzymatically released and procainamide-labeled N -glycans from human (blue, top) and canine (green, bottom) blood serum. ( B ) HILIC-UPLC profile of N -glycans released from canine IgG. IgG was purified using Protein G from canine serum. Glycan structures are annotated following the nomenclature outlined by the Consortium for Functional Glycomics (CFG). The inset in B shows the monosaccharide symbols.

    Journal: Glycobiology

    Article Title: Glycosylation profiling of dog serum reveals differences compared to human serum

    doi: 10.1093/glycob/cwy070

    Figure Lengend Snippet: ( A ) HILIC-UPLC profiles of enzymatically released and procainamide-labeled N -glycans from human (blue, top) and canine (green, bottom) blood serum. ( B ) HILIC-UPLC profile of N -glycans released from canine IgG. IgG was purified using Protein G from canine serum. Glycan structures are annotated following the nomenclature outlined by the Consortium for Functional Glycomics (CFG). The inset in B shows the monosaccharide symbols.

    Article Snippet: IgG was purified from canine serum using either Protein A or Protein G magnetic beads according to the manufacturers’ instructions (New England Biolabs).

    Techniques: Hydrophilic Interaction Liquid Chromatography, Labeling, Purification, Functional Assay