ph d 7 phage display peptide library kit  (New England Biolabs)


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    Name:
    Ph D 7 Phage Display Peptide Library Kit
    Description:
    Ph D 7 Phage Display Peptide Library Kit 10 panning exps
    Catalog Number:
    e8100s
    Price:
    554
    Size:
    10 exps
    Category:
    Phage Display Systems
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    Structured Review

    New England Biolabs ph d 7 phage display peptide library kit
    Ph D 7 Phage Display Peptide Library Kit
    Ph D 7 Phage Display Peptide Library Kit 10 panning exps
    https://www.bioz.com/result/ph d 7 phage display peptide library kit/product/New England Biolabs
    Average 94 stars, based on 68 article reviews
    Price from $9.99 to $1999.99
    ph d 7 phage display peptide library kit - by Bioz Stars, 2021-01
    94/100 stars

    Images

    1) Product Images from "Diagnostic Profiling of the Human Public IgM Repertoire With Scalable Mimotope Libraries"

    Article Title: Diagnostic Profiling of the Human Public IgM Repertoire With Scalable Mimotope Libraries

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2019.02796

    Distribution of the amino acid residues in the mimotope library. (A) Log odds (LO) relative to background frequencies. (B) Sequence logo of the LO by position relative to the overall background frequencies. (C) Sequence logo of the frequencies by position. (D) Sequence logo of the LO by position in an amplified Ph.D.-7 phage library without ligand selection relative to the overall background frequencies (based on W. L. Matochko, R. Derda. “Error analysis of deep sequencing of phage libraries: peptides censored in sequencing”. Comput Math Methods Med , 2013, 491612. 2013., http://www.chem.ualberta.ca/~derda/parasitepaper/rawfiles/PhD7-GAC-30FuR.txt ). (E) Sequence logo of the LO by position relative to the frequencies by position in the amplified, unselected library shown in (D) . The skewing of the distribution in the free N-terminus appears to be property of the library, whereas the selection by the IgM repertoire leads to slight skewing in the middle of the sequence toward negatively charged residues, glycine and tryptophan.
    Figure Legend Snippet: Distribution of the amino acid residues in the mimotope library. (A) Log odds (LO) relative to background frequencies. (B) Sequence logo of the LO by position relative to the overall background frequencies. (C) Sequence logo of the frequencies by position. (D) Sequence logo of the LO by position in an amplified Ph.D.-7 phage library without ligand selection relative to the overall background frequencies (based on W. L. Matochko, R. Derda. “Error analysis of deep sequencing of phage libraries: peptides censored in sequencing”. Comput Math Methods Med , 2013, 491612. 2013., http://www.chem.ualberta.ca/~derda/parasitepaper/rawfiles/PhD7-GAC-30FuR.txt ). (E) Sequence logo of the LO by position relative to the frequencies by position in the amplified, unselected library shown in (D) . The skewing of the distribution in the free N-terminus appears to be property of the library, whereas the selection by the IgM repertoire leads to slight skewing in the middle of the sequence toward negatively charged residues, glycine and tryptophan.

    Techniques Used: Sequencing, Amplification, Selection

    2) Product Images from "MRI-based molecular imaging of epicardium-derived stromal cells (EpiSC) by peptide-mediated active targeting"

    Article Title: MRI-based molecular imaging of epicardium-derived stromal cells (EpiSC) by peptide-mediated active targeting

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-78600-y

    ( a ) Schematic overview of the phage-display screening approach: phage from the PhD7 library were incubated with cultured EpiSCs, intensively washed and eluted at low pH (1st round of biopanning). Subsequently, isolated phage were incubated with blood to remove phage clones that bound to cellular components of blood (2nd round of biopanning). Phage isolated from the blood plasma were subjected to deep sequencing and bioinformatics analysis. ( b ) After incubation with the PhD7 phage library, EpiSCs were intensively washed and the amount of released phage per washing step was determined by qPCR (Mean ± SD; n = 5). ( c ) Total number of phage (± SD of n = 5) which were obtained after the first and second round of the biopanning procedure. ( d ) Enrichment of EpiSC specific peptides sequences (EP): the relative increase of the 50 most abundant peptide sequences in the five separate phage isolations from the 2nd round of the biopanning procedure. Enrichment was calculated in relation to the parent PhD7 phage library and is displayed as color-coded heat map. EPs (EP1-EP50) were arranged according to their mean relative enrichment. Prep = individual preparation number of EpiSCs. Parts of the figure were drawn by using pictures from Servier Medical Art ( http://smart.servier.com/ ).
    Figure Legend Snippet: ( a ) Schematic overview of the phage-display screening approach: phage from the PhD7 library were incubated with cultured EpiSCs, intensively washed and eluted at low pH (1st round of biopanning). Subsequently, isolated phage were incubated with blood to remove phage clones that bound to cellular components of blood (2nd round of biopanning). Phage isolated from the blood plasma were subjected to deep sequencing and bioinformatics analysis. ( b ) After incubation with the PhD7 phage library, EpiSCs were intensively washed and the amount of released phage per washing step was determined by qPCR (Mean ± SD; n = 5). ( c ) Total number of phage (± SD of n = 5) which were obtained after the first and second round of the biopanning procedure. ( d ) Enrichment of EpiSC specific peptides sequences (EP): the relative increase of the 50 most abundant peptide sequences in the five separate phage isolations from the 2nd round of the biopanning procedure. Enrichment was calculated in relation to the parent PhD7 phage library and is displayed as color-coded heat map. EPs (EP1-EP50) were arranged according to their mean relative enrichment. Prep = individual preparation number of EpiSCs. Parts of the figure were drawn by using pictures from Servier Medical Art ( http://smart.servier.com/ ).

    Techniques Used: Incubation, Cell Culture, Isolation, Clone Assay, Sequencing, Real-time Polymerase Chain Reaction

    3) Product Images from "A set of cancer stem cell homing peptides associating with the glycan moieties of glycosphingolipids"

    Article Title: A set of cancer stem cell homing peptides associating with the glycan moieties of glycosphingolipids

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24960

    Strategies for the establishment of primary and secondary CSC HP libraries (A) Flow chart of the primary screening from the original M13 PhD7 library to prepare the EMT6 CSC HP-Library was shown in the upper part. The secondary screening tactics to produce the CT26 CSC HP-Library, the Hepa1-6 CSC HP-Library, the CSC/ESC HP-Library, and the CSC HP-Library was illustrated in the lower part. (B) The same strategy of EMT6 CSC HP-library preparation was used to establish the PANC-1 CSC HP-library.
    Figure Legend Snippet: Strategies for the establishment of primary and secondary CSC HP libraries (A) Flow chart of the primary screening from the original M13 PhD7 library to prepare the EMT6 CSC HP-Library was shown in the upper part. The secondary screening tactics to produce the CT26 CSC HP-Library, the Hepa1-6 CSC HP-Library, the CSC/ESC HP-Library, and the CSC HP-Library was illustrated in the lower part. (B) The same strategy of EMT6 CSC HP-library preparation was used to establish the PANC-1 CSC HP-library.

    Techniques Used: Flow Cytometry

    4) Product Images from "Amsacta moorei Entomopoxvirus Inhibitor of Apoptosis Suppresses Cell Death by Binding Grim and Hid"

    Article Title: Amsacta moorei Entomopoxvirus Inhibitor of Apoptosis Suppresses Cell Death by Binding Grim and Hid

    Journal: Journal of Virology

    doi: 10.1128/JVI.79.6.3684-3691.2005

    Phage display sequences identified after three rounds of phage selection and amplification. (A) Twenty representative isolates from the Ph.D.-7 library (New England BioLabs) were sequenced after each round. Recurrences of specific amino acids after the third round are indicated in parentheses. X, no significant recurrence. (B) The binding motif sequences of mammalian (human) and insect ( Drosophila ) IBM-containing proteins. The first seven amino acids of the mature forms of each protein are shown. Superscripted numbers indicate the amino acid positions of the alanine in the precursor proteins.
    Figure Legend Snippet: Phage display sequences identified after three rounds of phage selection and amplification. (A) Twenty representative isolates from the Ph.D.-7 library (New England BioLabs) were sequenced after each round. Recurrences of specific amino acids after the third round are indicated in parentheses. X, no significant recurrence. (B) The binding motif sequences of mammalian (human) and insect ( Drosophila ) IBM-containing proteins. The first seven amino acids of the mature forms of each protein are shown. Superscripted numbers indicate the amino acid positions of the alanine in the precursor proteins.

    Techniques Used: Selection, Amplification, Binding Assay

    5) Product Images from "Mimotopes selected with neutralizing antibodies against multiple subtypes of influenza A"

    Article Title: Mimotopes selected with neutralizing antibodies against multiple subtypes of influenza A

    Journal: Virology Journal

    doi: 10.1186/1743-422X-8-542

    The ELISA results of top 12 phage clones with higher C179/BSA ELISA signal ratio from Ph.D. -7, Ph.D. -12 and Ph.D. -C7C peptide phage-display libraries .  a : Phage clones from Ph.D. -7 peptide phage-display library.  b : Phage clones from Ph.D. -12 peptide phage-display library;  c : Phage clones from Ph.D. -C7C peptide phage-display library.
    Figure Legend Snippet: The ELISA results of top 12 phage clones with higher C179/BSA ELISA signal ratio from Ph.D. -7, Ph.D. -12 and Ph.D. -C7C peptide phage-display libraries . a : Phage clones from Ph.D. -7 peptide phage-display library. b : Phage clones from Ph.D. -12 peptide phage-display library; c : Phage clones from Ph.D. -C7C peptide phage-display library.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Clone Assay

    6) Product Images from "Identification of Fusarium virguliforme FvTox1-Interacting Synthetic Peptides for Enhancing Foliar Sudden Death Syndrome Resistance in Soybean"

    Article Title: Identification of Fusarium virguliforme FvTox1-Interacting Synthetic Peptides for Enhancing Foliar Sudden Death Syndrome Resistance in Soybean

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0145156

    Diagrammatic representation of the workflow applied in affinity purification of M13 phage clones that displayed FvTox1-interacting synthetic peptides. (A), Bio-panning of the phage display libraries on plastic surface of a microtiter plates coated with 1.5 ml FvTox1 (30 ng/μl). Unbound phage particles were washed off; and M13 phage particles bound to FvTox1 were used to infect E . coli for starting a second round of panning. The process was repeated once more. (B), Plating of candidate M13 phage clones displaying FvTox1-interacting peptides. An eluate from the last panning in A was plated on X-gal/IPTG agar plates. (C), Identification of candidate M13 phage clones displaying FvTox1-interacting peptides. Phage clones were adsorbed onto nitrocellulose paper and hybridized to the His-tagged purified FvTox1 proteins. FvTox1-interacting clones were identified by detecting FvTox1 with the anti-His antibody. (D), Western blot analysis of the selected phage clones for interaction with FvTox1. Selected M13 phage particles from plates in C were transferred to nitrocellulose filters and hybridized to FvTox1, which was detected with an anti-His antibody. (E), Western blot analysis of the selected clones for interaction with FvTox1. Selected clones in D were reinvestigated for interaction with FvTox1, adsorbed onto a nitrocellulose membranes and detecting the interaction of individual clones to FvTox1 with an anti-M13 antibody (Details are presented in S2 Fig ). (F), Electropherogram of a nucleotide molecule encoding an FvTox1-intearcting peptide is presented.
    Figure Legend Snippet: Diagrammatic representation of the workflow applied in affinity purification of M13 phage clones that displayed FvTox1-interacting synthetic peptides. (A), Bio-panning of the phage display libraries on plastic surface of a microtiter plates coated with 1.5 ml FvTox1 (30 ng/μl). Unbound phage particles were washed off; and M13 phage particles bound to FvTox1 were used to infect E . coli for starting a second round of panning. The process was repeated once more. (B), Plating of candidate M13 phage clones displaying FvTox1-interacting peptides. An eluate from the last panning in A was plated on X-gal/IPTG agar plates. (C), Identification of candidate M13 phage clones displaying FvTox1-interacting peptides. Phage clones were adsorbed onto nitrocellulose paper and hybridized to the His-tagged purified FvTox1 proteins. FvTox1-interacting clones were identified by detecting FvTox1 with the anti-His antibody. (D), Western blot analysis of the selected phage clones for interaction with FvTox1. Selected M13 phage particles from plates in C were transferred to nitrocellulose filters and hybridized to FvTox1, which was detected with an anti-His antibody. (E), Western blot analysis of the selected clones for interaction with FvTox1. Selected clones in D were reinvestigated for interaction with FvTox1, adsorbed onto a nitrocellulose membranes and detecting the interaction of individual clones to FvTox1 with an anti-M13 antibody (Details are presented in S2 Fig ). (F), Electropherogram of a nucleotide molecule encoding an FvTox1-intearcting peptide is presented.

    Techniques Used: Affinity Purification, Clone Assay, Purification, Western Blot

    7) Product Images from "Peptide affinity reagents for AAV capsid recognition and purification"

    Article Title: Peptide affinity reagents for AAV capsid recognition and purification

    Journal: Gene therapy

    doi: 10.1038/gt.2011.46

    ( a ) Phage panning procedure. The Ph.D.-7 phage display library was co-incubated with AAV capsids to allow binding (B), followed by washing (W) of unbound phage and elution (E) of bound phage. Bound phage were then amplified and subjected to three rounds
    Figure Legend Snippet: ( a ) Phage panning procedure. The Ph.D.-7 phage display library was co-incubated with AAV capsids to allow binding (B), followed by washing (W) of unbound phage and elution (E) of bound phage. Bound phage were then amplified and subjected to three rounds

    Techniques Used: Incubation, Binding Assay, Amplification

    8) Product Images from "Synthetic Toll Like Receptor-4 (TLR-4) Agonist Peptides as a Novel Class of Adjuvants"

    Article Title: Synthetic Toll Like Receptor-4 (TLR-4) Agonist Peptides as a Novel Class of Adjuvants

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030839

    Identification of LPS specific peptide mimotopes. LPS antibody was immobilized at a concentration of 1 µg per well of a 96-well plate. Phages (2×10 11 ) expressing 7-mer peptides were initially added to the well of a 96-well plate containing immobilized LPS antibody after which non-specific phages were removed. Twelve random phage clones were selected from three rounds of panning and specificity to LPS antibody (black bars) was confirmed by ELISA using HSP70 antibody (white bars) as a negative control. All 12 (RS01-RS12) clones selected displayed specific reactivity to LPS antibody, as determined using the HRP labeled anti-M13 phage antibody which was detected by the HRP substrate SIGMAFAST™ OPD and absorbance measured at 490 nm. Each experiment was repeated three times with similar results observed and the standard deviation shown above each sample represents three replicates in one experiment.
    Figure Legend Snippet: Identification of LPS specific peptide mimotopes. LPS antibody was immobilized at a concentration of 1 µg per well of a 96-well plate. Phages (2×10 11 ) expressing 7-mer peptides were initially added to the well of a 96-well plate containing immobilized LPS antibody after which non-specific phages were removed. Twelve random phage clones were selected from three rounds of panning and specificity to LPS antibody (black bars) was confirmed by ELISA using HSP70 antibody (white bars) as a negative control. All 12 (RS01-RS12) clones selected displayed specific reactivity to LPS antibody, as determined using the HRP labeled anti-M13 phage antibody which was detected by the HRP substrate SIGMAFAST™ OPD and absorbance measured at 490 nm. Each experiment was repeated three times with similar results observed and the standard deviation shown above each sample represents three replicates in one experiment.

    Techniques Used: Concentration Assay, Expressing, Clone Assay, Enzyme-linked Immunosorbent Assay, Negative Control, Labeling, Standard Deviation

    9) Product Images from "Specific Targeting of Hepatitis C Virus Core Protein by an Intracellular Single-Chain Antibody of Human Origin"

    Article Title: Specific Targeting of Hepatitis C Virus Core Protein by an Intracellular Single-Chain Antibody of Human Origin

    Journal:

    doi: 10.1002/hep.22366

    Identification of scFv42C binding motif in HCV core protein by multiple sequence alignment. Core protein sequence (residues 79–88) is shown on the top, followed by deduced amino acid sequences of eight phage clones (mimotopes) from the Ph.D.-7
    Figure Legend Snippet: Identification of scFv42C binding motif in HCV core protein by multiple sequence alignment. Core protein sequence (residues 79–88) is shown on the top, followed by deduced amino acid sequences of eight phage clones (mimotopes) from the Ph.D.-7

    Techniques Used: Binding Assay, Sequencing

    Related Articles

    Produced:

    Article Title: Mimotopes selected with neutralizing antibodies against multiple subtypes of influenza A
    Article Snippet: .. Phage display libraries Ph.D.-7, Ph.D.-12 and Ph.D.-C7C were produced by New England Biolabs, Inc (Ipswich, MA, USA), with random linear 7-mer, 12-mer or cyclic 7-mer peptides fused to minor coat proteins (pIII) of M13 filamentous phages. ..

    Clone Assay:

    Article Title: MRI-based molecular imaging of epicardium-derived stromal cells (EpiSC) by peptide-mediated active targeting
    Article Snippet: .. Biopanning of a phage library on cultivated EpiSCs and whole blood A PhD7 phage library (109 different phage clones; New England Biolabs GmbH, Frankfurt am Main, Germany) was used for the biopanning procedure on EpiSCs which had been seeded 48 h before. .. 5 × 1010 plaque forming units (PFU) were incubated for 1.5 h at 37 °C in a volume of 1 ml cell culture medium on 3 × 105 EpiSCs.

    Library Screening:

    Article Title: Identification of Fusarium virguliforme FvTox1-Interacting Synthetic Peptides for Enhancing Foliar Sudden Death Syndrome Resistance in Soybean
    Article Snippet: .. Phage Display Peptide Library Screening Three phage display peptides libraries, Ph.D.-7, Ph.D.-12 and Ph.D-C7C were obtained from NEB Inc. (New England Lab, Woburn, MA). .. FvTox1 was expressed in an insect cell line and immobilized on a plate for screening the phage display libraries using a modified NEB, Inc. screening protocol ([ ]; ).

    Expressing:

    Article Title: Amsacta moorei Entomopoxvirus Inhibitor of Apoptosis Suppresses Cell Death by Binding Grim and Hid
    Article Snippet: .. A phage library (Ph.D.-7 Phage Display Peptide Library kit; New England BioLabs) expressing seven random amino acids fused to the amino terminus of the mature PIII coat protein was used for screening. .. For each GST-BIR protein, three sequential round of affinity purification and reamplification of the phage were carried out, with ≥20 phage isolates sequenced after each round of selection.

    Concentration Assay:

    Article Title: Diagnostic Profiling of the Human Public IgM Repertoire With Scalable Mimotope Libraries
    Article Snippet: .. A 7-mer random peptide library (E8100S, Ph.D.-7, New England Biolabs, USA) was panned overnight at 4°C on pooled human IgM adsorbed on polystyrene plates at a concentration of 0.1 mg/ml, washed, eluted with glycine buffer at pH 2.7, and immediately brought to pH 7. .. The eluate was transferred to a plate coated with monoclonal IgM and incubated according to the same protocol, but this time, the phage solution was collected after adsorption and amplified once, following the procedure described by Matochko et al. ( ).

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    New England Biolabs ph d 7 phage display peptide library kit
    Distribution of the amino acid residues in the mimotope library. (A) Log odds (LO) relative to background frequencies. (B) Sequence logo of the LO by position relative to the overall background frequencies. (C) Sequence logo of the frequencies by position. (D) Sequence logo of the LO by position in an amplified <t>Ph.D.-7</t> phage library without ligand selection relative to the overall background frequencies (based on W. L. Matochko, R. Derda. “Error analysis of deep sequencing of phage libraries: peptides censored in sequencing”. Comput Math Methods Med , 2013, 491612. 2013., http://www.chem.ualberta.ca/~derda/parasitepaper/rawfiles/PhD7-GAC-30FuR.txt ). (E) Sequence logo of the LO by position relative to the frequencies by position in the amplified, unselected library shown in (D) . The skewing of the distribution in the free N-terminus appears to be property of the library, whereas the selection by the IgM repertoire leads to slight skewing in the middle of the sequence toward negatively charged residues, glycine and tryptophan.
    Ph D 7 Phage Display Peptide Library Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ph d 7 phage display peptide library kit/product/New England Biolabs
    Average 94 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    ph d 7 phage display peptide library kit - by Bioz Stars, 2021-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    Distribution of the amino acid residues in the mimotope library. (A) Log odds (LO) relative to background frequencies. (B) Sequence logo of the LO by position relative to the overall background frequencies. (C) Sequence logo of the frequencies by position. (D) Sequence logo of the LO by position in an amplified Ph.D.-7 phage library without ligand selection relative to the overall background frequencies (based on W. L. Matochko, R. Derda. “Error analysis of deep sequencing of phage libraries: peptides censored in sequencing”. Comput Math Methods Med , 2013, 491612. 2013., http://www.chem.ualberta.ca/~derda/parasitepaper/rawfiles/PhD7-GAC-30FuR.txt ). (E) Sequence logo of the LO by position relative to the frequencies by position in the amplified, unselected library shown in (D) . The skewing of the distribution in the free N-terminus appears to be property of the library, whereas the selection by the IgM repertoire leads to slight skewing in the middle of the sequence toward negatively charged residues, glycine and tryptophan.

    Journal: Frontiers in Immunology

    Article Title: Diagnostic Profiling of the Human Public IgM Repertoire With Scalable Mimotope Libraries

    doi: 10.3389/fimmu.2019.02796

    Figure Lengend Snippet: Distribution of the amino acid residues in the mimotope library. (A) Log odds (LO) relative to background frequencies. (B) Sequence logo of the LO by position relative to the overall background frequencies. (C) Sequence logo of the frequencies by position. (D) Sequence logo of the LO by position in an amplified Ph.D.-7 phage library without ligand selection relative to the overall background frequencies (based on W. L. Matochko, R. Derda. “Error analysis of deep sequencing of phage libraries: peptides censored in sequencing”. Comput Math Methods Med , 2013, 491612. 2013., http://www.chem.ualberta.ca/~derda/parasitepaper/rawfiles/PhD7-GAC-30FuR.txt ). (E) Sequence logo of the LO by position relative to the frequencies by position in the amplified, unselected library shown in (D) . The skewing of the distribution in the free N-terminus appears to be property of the library, whereas the selection by the IgM repertoire leads to slight skewing in the middle of the sequence toward negatively charged residues, glycine and tryptophan.

    Article Snippet: A 7-mer random peptide library (E8100S, Ph.D.-7, New England Biolabs, USA) was panned overnight at 4°C on pooled human IgM adsorbed on polystyrene plates at a concentration of 0.1 mg/ml, washed, eluted with glycine buffer at pH 2.7, and immediately brought to pH 7.

    Techniques: Sequencing, Amplification, Selection

    ( a ) Schematic overview of the phage-display screening approach: phage from the PhD7 library were incubated with cultured EpiSCs, intensively washed and eluted at low pH (1st round of biopanning). Subsequently, isolated phage were incubated with blood to remove phage clones that bound to cellular components of blood (2nd round of biopanning). Phage isolated from the blood plasma were subjected to deep sequencing and bioinformatics analysis. ( b ) After incubation with the PhD7 phage library, EpiSCs were intensively washed and the amount of released phage per washing step was determined by qPCR (Mean ± SD; n = 5). ( c ) Total number of phage (± SD of n = 5) which were obtained after the first and second round of the biopanning procedure. ( d ) Enrichment of EpiSC specific peptides sequences (EP): the relative increase of the 50 most abundant peptide sequences in the five separate phage isolations from the 2nd round of the biopanning procedure. Enrichment was calculated in relation to the parent PhD7 phage library and is displayed as color-coded heat map. EPs (EP1-EP50) were arranged according to their mean relative enrichment. Prep = individual preparation number of EpiSCs. Parts of the figure were drawn by using pictures from Servier Medical Art ( http://smart.servier.com/ ).

    Journal: Scientific Reports

    Article Title: MRI-based molecular imaging of epicardium-derived stromal cells (EpiSC) by peptide-mediated active targeting

    doi: 10.1038/s41598-020-78600-y

    Figure Lengend Snippet: ( a ) Schematic overview of the phage-display screening approach: phage from the PhD7 library were incubated with cultured EpiSCs, intensively washed and eluted at low pH (1st round of biopanning). Subsequently, isolated phage were incubated with blood to remove phage clones that bound to cellular components of blood (2nd round of biopanning). Phage isolated from the blood plasma were subjected to deep sequencing and bioinformatics analysis. ( b ) After incubation with the PhD7 phage library, EpiSCs were intensively washed and the amount of released phage per washing step was determined by qPCR (Mean ± SD; n = 5). ( c ) Total number of phage (± SD of n = 5) which were obtained after the first and second round of the biopanning procedure. ( d ) Enrichment of EpiSC specific peptides sequences (EP): the relative increase of the 50 most abundant peptide sequences in the five separate phage isolations from the 2nd round of the biopanning procedure. Enrichment was calculated in relation to the parent PhD7 phage library and is displayed as color-coded heat map. EPs (EP1-EP50) were arranged according to their mean relative enrichment. Prep = individual preparation number of EpiSCs. Parts of the figure were drawn by using pictures from Servier Medical Art ( http://smart.servier.com/ ).

    Article Snippet: Biopanning of a phage library on cultivated EpiSCs and whole blood A PhD7 phage library (109 different phage clones; New England Biolabs GmbH, Frankfurt am Main, Germany) was used for the biopanning procedure on EpiSCs which had been seeded 48 h before.

    Techniques: Incubation, Cell Culture, Isolation, Clone Assay, Sequencing, Real-time Polymerase Chain Reaction

    Phage display sequences identified after three rounds of phage selection and amplification. (A) Twenty representative isolates from the Ph.D.-7 library (New England BioLabs) were sequenced after each round. Recurrences of specific amino acids after the third round are indicated in parentheses. X, no significant recurrence. (B) The binding motif sequences of mammalian (human) and insect ( Drosophila ) IBM-containing proteins. The first seven amino acids of the mature forms of each protein are shown. Superscripted numbers indicate the amino acid positions of the alanine in the precursor proteins.

    Journal: Journal of Virology

    Article Title: Amsacta moorei Entomopoxvirus Inhibitor of Apoptosis Suppresses Cell Death by Binding Grim and Hid

    doi: 10.1128/JVI.79.6.3684-3691.2005

    Figure Lengend Snippet: Phage display sequences identified after three rounds of phage selection and amplification. (A) Twenty representative isolates from the Ph.D.-7 library (New England BioLabs) were sequenced after each round. Recurrences of specific amino acids after the third round are indicated in parentheses. X, no significant recurrence. (B) The binding motif sequences of mammalian (human) and insect ( Drosophila ) IBM-containing proteins. The first seven amino acids of the mature forms of each protein are shown. Superscripted numbers indicate the amino acid positions of the alanine in the precursor proteins.

    Article Snippet: A phage library (Ph.D.-7 Phage Display Peptide Library kit; New England BioLabs) expressing seven random amino acids fused to the amino terminus of the mature PIII coat protein was used for screening.

    Techniques: Selection, Amplification, Binding Assay

    The ELISA results of top 12 phage clones with higher C179/BSA ELISA signal ratio from Ph.D. -7, Ph.D. -12 and Ph.D. -C7C peptide phage-display libraries .  a : Phage clones from Ph.D. -7 peptide phage-display library.  b : Phage clones from Ph.D. -12 peptide phage-display library;  c : Phage clones from Ph.D. -C7C peptide phage-display library.

    Journal: Virology Journal

    Article Title: Mimotopes selected with neutralizing antibodies against multiple subtypes of influenza A

    doi: 10.1186/1743-422X-8-542

    Figure Lengend Snippet: The ELISA results of top 12 phage clones with higher C179/BSA ELISA signal ratio from Ph.D. -7, Ph.D. -12 and Ph.D. -C7C peptide phage-display libraries . a : Phage clones from Ph.D. -7 peptide phage-display library. b : Phage clones from Ph.D. -12 peptide phage-display library; c : Phage clones from Ph.D. -C7C peptide phage-display library.

    Article Snippet: Phage display libraries Ph.D.-7, Ph.D.-12 and Ph.D.-C7C were produced by New England Biolabs, Inc (Ipswich, MA, USA), with random linear 7-mer, 12-mer or cyclic 7-mer peptides fused to minor coat proteins (pIII) of M13 filamentous phages.

    Techniques: Enzyme-linked Immunosorbent Assay, Clone Assay