m-280 streptavidin coated dynabeads Search Results


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    Thermo Fisher streptavidin coated m280 dynabeads
    Histone fate after in vitro chromatin resection by the Sgs1-Dna2 pathway. A , native PAGE of a 500-bp DNA fragment harboring a central, 601-positioning sequence reconstituted into mononucleosomes by salt step dialysis at different ratios ( r ) of histone octamers to DNA. Note that the minor nucleosome species is likely to represent a nucleosome assembled on the DNA end. B and C , chromatin resection time course with 3′-radiolabeled chromatin and reactions that contain Mre11-Rad50-Xrs2, Sgs1, Top3-Rmi complex, Dna2, and RPA. C, addition of <t>streptavidin-coated</t> magnetic beads inhibit chromatin resection on one strand. Note the appearance of slower migrating ssDNA. D, analysis of DNA and protein content following magnetic DNA pulldown after chromatin resection. Left panel , radiolabel analysis of DNA before (−) and after (+) resection. One sample was also treated with EcoRI prior to resection and the released DNA was analyzed. Right panel , histone immunoblotting of bead bound ( B ) and unbound ( U ) fractions before and after chromatin resection.
    Streptavidin Coated M280 Dynabeads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin coated m280 dynabeads/product/Thermo Fisher
    Average 99 stars, based on 156 article reviews
    Price from $9.99 to $1999.99
    streptavidin coated m280 dynabeads - by Bioz Stars, 2020-05
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    95
    Thermo Fisher streptavidin coated magnetic beads
    Binding specificities of anti-VEGF-C scFv. (A) ELISA screening of random clones obtained after 2 or 3 rounds of panning against ΔNΔC-VEGF-C. (B) ELISA analysis of representative anti-VEGF-C scFv clones for the 4 different amino acid sequences obtained. Maxisorp or <t>streptavidin-precoated</t> (SA) plates were coated with his-tagged human ΔNΔC-VEGF-C derived from P. pastoris or biotinylated his-tagged human ΔNΔC-VEGF-C from mammalian cells or P. pastoris , respectively. Control surfaces were left untreated. Antibody fragments and control antibodies were subsequently added and the ELISA was developed as described in Materials and Methods. (C) Cross-reactivity tested by ELISA. Human ΔNΔC-VEGF-C orΔNΔC-VEGF-D (both from mammalian cells) were coated on a maxisorp plate. Anti-VEGF-C scFv clone VC2 or a negative control (PBS only) was added and the ELISA was developed as described in Materials and Methods. (D) BIAcore profiles from the 4 different anti-VEGF-C scFv clones. Different concentrations of protein-A purified scFv were injected on a streptavidin-precoated sensorchip coated with ca. 2000 RU biotinylated mammalian cell-derived ΔNΔC-VEGF-C.
    Streptavidin Coated Magnetic Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 4182 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin coated magnetic beads/product/Thermo Fisher
    Average 95 stars, based on 4182 article reviews
    Price from $9.99 to $1999.99
    streptavidin coated magnetic beads - by Bioz Stars, 2020-05
    95/100 stars
      Buy from Supplier

    Image Search Results


    Histone fate after in vitro chromatin resection by the Sgs1-Dna2 pathway. A , native PAGE of a 500-bp DNA fragment harboring a central, 601-positioning sequence reconstituted into mononucleosomes by salt step dialysis at different ratios ( r ) of histone octamers to DNA. Note that the minor nucleosome species is likely to represent a nucleosome assembled on the DNA end. B and C , chromatin resection time course with 3′-radiolabeled chromatin and reactions that contain Mre11-Rad50-Xrs2, Sgs1, Top3-Rmi complex, Dna2, and RPA. C, addition of streptavidin-coated magnetic beads inhibit chromatin resection on one strand. Note the appearance of slower migrating ssDNA. D, analysis of DNA and protein content following magnetic DNA pulldown after chromatin resection. Left panel , radiolabel analysis of DNA before (−) and after (+) resection. One sample was also treated with EcoRI prior to resection and the released DNA was analyzed. Right panel , histone immunoblotting of bead bound ( B ) and unbound ( U ) fractions before and after chromatin resection.

    Journal: The Journal of Biological Chemistry

    Article Title: Nucleosome-like, Single-stranded DNA (ssDNA)-Histone Octamer Complexes and the Implication for DNA Double Strand Break Repair *

    doi: 10.1074/jbc.M117.776369

    Figure Lengend Snippet: Histone fate after in vitro chromatin resection by the Sgs1-Dna2 pathway. A , native PAGE of a 500-bp DNA fragment harboring a central, 601-positioning sequence reconstituted into mononucleosomes by salt step dialysis at different ratios ( r ) of histone octamers to DNA. Note that the minor nucleosome species is likely to represent a nucleosome assembled on the DNA end. B and C , chromatin resection time course with 3′-radiolabeled chromatin and reactions that contain Mre11-Rad50-Xrs2, Sgs1, Top3-Rmi complex, Dna2, and RPA. C, addition of streptavidin-coated magnetic beads inhibit chromatin resection on one strand. Note the appearance of slower migrating ssDNA. D, analysis of DNA and protein content following magnetic DNA pulldown after chromatin resection. Left panel , radiolabel analysis of DNA before (−) and after (+) resection. One sample was also treated with EcoRI prior to resection and the released DNA was analyzed. Right panel , histone immunoblotting of bead bound ( B ) and unbound ( U ) fractions before and after chromatin resection.

    Article Snippet: Chromatin was magnetically precipitated with streptavidin-coated beads (Dynabeads M-280 Streptavidin, Invitrogen) and beads were then washed twice with 10 m m Tris-HCl, pH 7.5, 50 m m NaCl, 0.5 mg/ml of BSA to remove unbound chromatin.

    Techniques: In Vitro, Clear Native PAGE, Sequencing, Recombinase Polymerase Amplification, Magnetic Beads

    Biochemical characterization of reconstituted mononucleosomes. A, native PAGE of the indicated nucleic acid with increasing histone ratios ( r ) after reconstitution by salt step dialysis. Note that ssDNA stains less intensely with ethidium bromide. B , 4% native PAGE of ssDNA-histone octamer reconstitutions, using radiolabeled DNA fragments of varying length. r , histone octamer:DNA molar ratio. C, the stability of double-stranded and single-stranded nucleosomes after a 1-h incubation at the indicated range of temperatures, analyzed by separation on a 4% native PAGE. D, 200-nt ssDNA or 200-bp dsDNA chromatin reconstitutions were immobilized on streptavidin-coated magnetic beads, and histone content was analyzed after magnetic pulldown by SDS-PAGE. E , the indicated histone complexes were used in chromatin reconstitution reactions with 150-bp or 150-nt DNA fragments. Reconstitutions were analyzed by 4% native PAGE.

    Journal: The Journal of Biological Chemistry

    Article Title: Nucleosome-like, Single-stranded DNA (ssDNA)-Histone Octamer Complexes and the Implication for DNA Double Strand Break Repair *

    doi: 10.1074/jbc.M117.776369

    Figure Lengend Snippet: Biochemical characterization of reconstituted mononucleosomes. A, native PAGE of the indicated nucleic acid with increasing histone ratios ( r ) after reconstitution by salt step dialysis. Note that ssDNA stains less intensely with ethidium bromide. B , 4% native PAGE of ssDNA-histone octamer reconstitutions, using radiolabeled DNA fragments of varying length. r , histone octamer:DNA molar ratio. C, the stability of double-stranded and single-stranded nucleosomes after a 1-h incubation at the indicated range of temperatures, analyzed by separation on a 4% native PAGE. D, 200-nt ssDNA or 200-bp dsDNA chromatin reconstitutions were immobilized on streptavidin-coated magnetic beads, and histone content was analyzed after magnetic pulldown by SDS-PAGE. E , the indicated histone complexes were used in chromatin reconstitution reactions with 150-bp or 150-nt DNA fragments. Reconstitutions were analyzed by 4% native PAGE.

    Article Snippet: Chromatin was magnetically precipitated with streptavidin-coated beads (Dynabeads M-280 Streptavidin, Invitrogen) and beads were then washed twice with 10 m m Tris-HCl, pH 7.5, 50 m m NaCl, 0.5 mg/ml of BSA to remove unbound chromatin.

    Techniques: Clear Native PAGE, Incubation, Magnetic Beads, SDS Page

    DSBs enrichment workflow and specificity of the DNA DSBs ( A ) DSBs enrichment workflow by MTX treatment or Restriction endonuclease digestion for quality control. Fragments released from the streptavidin beads were amplified by PCR using sequencing primers and sequenced. ( B ) Quality control of in situ digestion and blunt-ending by capillary electrophoresis. The top two traces are for the endonuclease digestion and blunt-ending performed in liquid; the bottom two traces are in low melting point agarose gel. I represents the size of digestion product of a 567-bp fluorescence-labeled DNA fragment by restriction digestion while II shows the size of digestion product after blunt-ending; III and IV represent the above reactions respectively in low melting point agarose gel. The arrow in black represents the complete blunt-ending. X-axis represents the size of fragments(bp), Y-axis represents the detector signal of peak(rfu). ( C ) DSBs enrichment products separated by agarose gel electrophoresis indicated by white box. ( D–F ) Capillary electrophoresis to detect DSB enrichment products after SbfI ( D ), PmeI ( E ) and HindIII ( F ) digestion. TA clone sequencing confirmed the results. The arrow in red indicates the DSB enrichments on Capillary electrophoresis; the circle marked with red-dotted lines shows the restriction sites; the arrow in black shows the ligation point. X-axis represents the size of fragments(bp), while Y-axis represents the detector signal of peak(rfu). ( G, H ) Capillary electrophoresis to detect DSB enrichment products of normal mESCs cultured in complete medium ( G ) and cultured in complete medium with 0.12 μM MTX ( H ). The arrow in red indicates the DSB enrichments. X-axis represents the size of fragments(bp), while Y-axis represents the detector signal of peak(rfu).

    Journal: Nucleic Acids Research

    Article Title: Folate deficiency facilitates recruitment of upstream binding factor to hot spots of DNA double-strand breaks of rRNA genes and promotes its transcription

    doi: 10.1093/nar/gkw1208

    Figure Lengend Snippet: DSBs enrichment workflow and specificity of the DNA DSBs ( A ) DSBs enrichment workflow by MTX treatment or Restriction endonuclease digestion for quality control. Fragments released from the streptavidin beads were amplified by PCR using sequencing primers and sequenced. ( B ) Quality control of in situ digestion and blunt-ending by capillary electrophoresis. The top two traces are for the endonuclease digestion and blunt-ending performed in liquid; the bottom two traces are in low melting point agarose gel. I represents the size of digestion product of a 567-bp fluorescence-labeled DNA fragment by restriction digestion while II shows the size of digestion product after blunt-ending; III and IV represent the above reactions respectively in low melting point agarose gel. The arrow in black represents the complete blunt-ending. X-axis represents the size of fragments(bp), Y-axis represents the detector signal of peak(rfu). ( C ) DSBs enrichment products separated by agarose gel electrophoresis indicated by white box. ( D–F ) Capillary electrophoresis to detect DSB enrichment products after SbfI ( D ), PmeI ( E ) and HindIII ( F ) digestion. TA clone sequencing confirmed the results. The arrow in red indicates the DSB enrichments on Capillary electrophoresis; the circle marked with red-dotted lines shows the restriction sites; the arrow in black shows the ligation point. X-axis represents the size of fragments(bp), while Y-axis represents the detector signal of peak(rfu). ( G, H ) Capillary electrophoresis to detect DSB enrichment products of normal mESCs cultured in complete medium ( G ) and cultured in complete medium with 0.12 μM MTX ( H ). The arrow in red indicates the DSB enrichments. X-axis represents the size of fragments(bp), while Y-axis represents the detector signal of peak(rfu).

    Article Snippet: The biotinylated ends were captured with 50 μl of streptavidin-coated paramagnetic beads (Dynabeads® M-280 Streptavidin, Invitrogen), and the DNA was then ligated to 60 pmol linker 2 in a 100 μl reaction volume containing 400 U of T4 DNA ligase by incubation at 16°C for 4 h. The precipitate was gently resuspended in 100 μl of 0.15 M NaOH at room temperature for 10 min and eluted with 30 μl 10 mM Tris–Cl (pH 8.0).

    Techniques: Amplification, Polymerase Chain Reaction, Sequencing, In Situ, Electrophoresis, Agarose Gel Electrophoresis, Fluorescence, Labeling, Ligation, Cell Culture

    Components and performance of the INTACT system ( A ) Confocal projection of the differentiation zone of an ADF8p : NTF/ACT2p : BirA transgenic root showing expression of the NTF in hair cells. GFP signal is shown in green and propidium iodide staining of cell walls is shown in red. ( B ) Confocal projection of the differentiation zone of an GL2p : NTF/ACT2p : BirA transgenic root showing expression of the NTF in non-hair cells. ( C ) Confocal section of the post-meristematic region of a GL2p : NTF/ACT2p : BirA transgenic root. ( D ) Fluorescence micrograph of nuclei (one is shown in inset) isolated from ADF8p : NTF/ACT2p : BirA transgenic roots and incubated with streptavidin Dynabeads. GFP and beads are shown in green and DAPI staining of DNA is shown in blue. ( E ) Streptavidin western blot of whole cell extracts (input) and anti-GFP immunoprecipitates (IP) from roots of ACT2p : BirA, ADF8p : NTF/ACT2p : BirA , and GL2p : NTF/ACT2p : BirA plants. Top and bottom bands in each lane are endogenous biotinylated proteins and the middle band is the 42 kD NTF. ( F .

    Journal: Developmental cell

    Article Title: A simple method for gene expression and chromatin profiling of individual cell types within a tissue

    doi: 10.1016/j.devcel.2010.05.013

    Figure Lengend Snippet: Components and performance of the INTACT system ( A ) Confocal projection of the differentiation zone of an ADF8p : NTF/ACT2p : BirA transgenic root showing expression of the NTF in hair cells. GFP signal is shown in green and propidium iodide staining of cell walls is shown in red. ( B ) Confocal projection of the differentiation zone of an GL2p : NTF/ACT2p : BirA transgenic root showing expression of the NTF in non-hair cells. ( C ) Confocal section of the post-meristematic region of a GL2p : NTF/ACT2p : BirA transgenic root. ( D ) Fluorescence micrograph of nuclei (one is shown in inset) isolated from ADF8p : NTF/ACT2p : BirA transgenic roots and incubated with streptavidin Dynabeads. GFP and beads are shown in green and DAPI staining of DNA is shown in blue. ( E ) Streptavidin western blot of whole cell extracts (input) and anti-GFP immunoprecipitates (IP) from roots of ACT2p : BirA, ADF8p : NTF/ACT2p : BirA , and GL2p : NTF/ACT2p : BirA plants. Top and bottom bands in each lane are endogenous biotinylated proteins and the middle band is the 42 kD NTF. ( F .

    Article Snippet: Twenty-five microliters of Invitrogen M-280 streptavidin-coated dynabeads (~1.5 × 107 beads) were added to the nuclear suspensions and this mixture was rotated at 4° C for 30 min to allow binding of beads to the biotinylated nuclei.

    Techniques: Transgenic Assay, Expressing, Staining, Fluorescence, Isolation, Incubation, Western Blot

    Schematic drawing of the PrASE procedures. A nested multiplex PCR amplification is performed with biotinylated inner PCR primers, generating short PCR products. The biotinylated PCR products are immobilized to streptavidin-coated magnetic beads, and ssDNA is generated by alkali elution of the non-biotinylated strand followed by annealing of ASE primers to the immobilized DNA strands. A washing step, where unannealed extension primers are removed, is then followed by PrASE with Cy5-labeled dCTPs and dUTPs. The last steps involve removal of unreacted nucleotides and PrASE enzymes, release of PrASE products by alkali elution and hybridization to tag microarrays. All procedures except the last step of hybridization to tag arrays are automated.

    Journal: Nucleic Acids Research

    Article Title: Competitive enzymatic reaction to control allele-specific extensions

    doi: 10.1093/nar/gni048

    Figure Lengend Snippet: Schematic drawing of the PrASE procedures. A nested multiplex PCR amplification is performed with biotinylated inner PCR primers, generating short PCR products. The biotinylated PCR products are immobilized to streptavidin-coated magnetic beads, and ssDNA is generated by alkali elution of the non-biotinylated strand followed by annealing of ASE primers to the immobilized DNA strands. A washing step, where unannealed extension primers are removed, is then followed by PrASE with Cy5-labeled dCTPs and dUTPs. The last steps involve removal of unreacted nucleotides and PrASE enzymes, release of PrASE products by alkali elution and hybridization to tag microarrays. All procedures except the last step of hybridization to tag arrays are automated.

    Article Snippet: An aliquot of 200 μg streptavidin-coated super paramagnetic beads (Dynabeads M280; Dynal, Oslo, Norway) were used to immobilize each multiplexed inner PCR product.

    Techniques: Multiplex Assay, Polymerase Chain Reaction, Amplification, Magnetic Beads, Generated, Labeling, Hybridization

    GNFRYLAPP inhibits DNA-PKcs in cellulo . A, Biotinylated GNFRYLAPP-pen, detected with FITC-streptavidin, formed foci in DNA-PKcs +/+ cells (KS1767) 1 h after 3 Gy of radiation, but did not form foci in DNA-PK -/- cells (M059J) treated the same way (left). Quantification of GNFRYLAPP-pen foci in irradiated KS1767 cells (3 Gy) reveals slow resolution of these foci over time (right). B, Phosphorylated DNA-PKcs (T2609) was detected 30 minutes after 3 Gy of radiation in irradiated KS1767 cells treated with scrambled peptide, but no signal was detected in the presence of GNFRYLAPP-pen. C, Two BRCA-deficient cell lines (HCC1937, BRCA1-null; CAPAN1, BRCA2-null) showed increased residual double-strand breaks by the neutral comet assay (see Materials and Methods) at 3 and 6 hours after 6 Gy of radiation when treated with GNFRYLAPP-pen (red) or with a DNA-PKcs active site inhibitor (DMNB, orange). D-E, Both of these lines also exhibited increased radiosensitivity by clonogenic assay (see Materials and Methods) when exposed to GNFRYLAPP-pen (red) or DMNB (orange). All peptides and small molecule inhibitors were used at 10 μM.

    Journal: Cancer research

    Article Title: Discovery of DNA repair inhibitors by combinatorial library profiling

    doi: 10.1158/0008-5472.CAN-10-2361

    Figure Lengend Snippet: GNFRYLAPP inhibits DNA-PKcs in cellulo . A, Biotinylated GNFRYLAPP-pen, detected with FITC-streptavidin, formed foci in DNA-PKcs +/+ cells (KS1767) 1 h after 3 Gy of radiation, but did not form foci in DNA-PK -/- cells (M059J) treated the same way (left). Quantification of GNFRYLAPP-pen foci in irradiated KS1767 cells (3 Gy) reveals slow resolution of these foci over time (right). B, Phosphorylated DNA-PKcs (T2609) was detected 30 minutes after 3 Gy of radiation in irradiated KS1767 cells treated with scrambled peptide, but no signal was detected in the presence of GNFRYLAPP-pen. C, Two BRCA-deficient cell lines (HCC1937, BRCA1-null; CAPAN1, BRCA2-null) showed increased residual double-strand breaks by the neutral comet assay (see Materials and Methods) at 3 and 6 hours after 6 Gy of radiation when treated with GNFRYLAPP-pen (red) or with a DNA-PKcs active site inhibitor (DMNB, orange). D-E, Both of these lines also exhibited increased radiosensitivity by clonogenic assay (see Materials and Methods) when exposed to GNFRYLAPP-pen (red) or DMNB (orange). All peptides and small molecule inhibitors were used at 10 μM.

    Article Snippet: A phage random peptide library displaying the insert XXXXYXXXX (X, any residue; Y, tyrosine) was generated. pSP72 plasmid DNA (Promega) was linearized, biotinylated, and bound to magnetic streptavidin-coated beads (Dynabeads M-280, Invitrogen) at a DNA:bead molar ratio of 60,000:1.

    Techniques: Irradiation, Neutral Comet Assay, Clonogenic Assay

    Binding specificities of anti-VEGF-C scFv. (A) ELISA screening of random clones obtained after 2 or 3 rounds of panning against ΔNΔC-VEGF-C. (B) ELISA analysis of representative anti-VEGF-C scFv clones for the 4 different amino acid sequences obtained. Maxisorp or streptavidin-precoated (SA) plates were coated with his-tagged human ΔNΔC-VEGF-C derived from P. pastoris or biotinylated his-tagged human ΔNΔC-VEGF-C from mammalian cells or P. pastoris , respectively. Control surfaces were left untreated. Antibody fragments and control antibodies were subsequently added and the ELISA was developed as described in Materials and Methods. (C) Cross-reactivity tested by ELISA. Human ΔNΔC-VEGF-C orΔNΔC-VEGF-D (both from mammalian cells) were coated on a maxisorp plate. Anti-VEGF-C scFv clone VC2 or a negative control (PBS only) was added and the ELISA was developed as described in Materials and Methods. (D) BIAcore profiles from the 4 different anti-VEGF-C scFv clones. Different concentrations of protein-A purified scFv were injected on a streptavidin-precoated sensorchip coated with ca. 2000 RU biotinylated mammalian cell-derived ΔNΔC-VEGF-C.

    Journal: PLoS ONE

    Article Title: Phage-Derived Fully Human Monoclonal Antibody Fragments to Human Vascular Endothelial Growth Factor-C Block Its Interaction with VEGF Receptor-2 and 3

    doi: 10.1371/journal.pone.0011941

    Figure Lengend Snippet: Binding specificities of anti-VEGF-C scFv. (A) ELISA screening of random clones obtained after 2 or 3 rounds of panning against ΔNΔC-VEGF-C. (B) ELISA analysis of representative anti-VEGF-C scFv clones for the 4 different amino acid sequences obtained. Maxisorp or streptavidin-precoated (SA) plates were coated with his-tagged human ΔNΔC-VEGF-C derived from P. pastoris or biotinylated his-tagged human ΔNΔC-VEGF-C from mammalian cells or P. pastoris , respectively. Control surfaces were left untreated. Antibody fragments and control antibodies were subsequently added and the ELISA was developed as described in Materials and Methods. (C) Cross-reactivity tested by ELISA. Human ΔNΔC-VEGF-C orΔNΔC-VEGF-D (both from mammalian cells) were coated on a maxisorp plate. Anti-VEGF-C scFv clone VC2 or a negative control (PBS only) was added and the ELISA was developed as described in Materials and Methods. (D) BIAcore profiles from the 4 different anti-VEGF-C scFv clones. Different concentrations of protein-A purified scFv were injected on a streptavidin-precoated sensorchip coated with ca. 2000 RU biotinylated mammalian cell-derived ΔNΔC-VEGF-C.

    Article Snippet: Prior to selection, 50 µl streptavidin coated magnetic beads (M-280 Dynabeads, Dynal Biotech, Oslo, Norway) per selection were blocked with 1 ml 3% BSA in PBS for 1 hour at room temperature and resuspended in 50 µl 2% BSA/PBS.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Clone Assay, Derivative Assay, Negative Control, Purification, Injection

    Affinity matured anti-VEGF-C scFvs possess a higher affinity. (A, B) ELISA analysis of bacterial supernatant from randomly picked affinity matured clones after 1 to 3 rounds of selection on biotinylated (A) P. pastoris -derived or (B) mammalian cell-derived ΔNΔC-VEGF-C. (C) BIAcore profiles of monomeric affinity matured anti-VEGF-C scFvs. Monomeric fractions of protein-A purified scFv were prepared by FPLC and injected as 2-fold dilution series on a streptavidin-sensorchip coated with 2000 RU biotinylated ΔNΔC-VEGF-C derived from mammalian cells.

    Journal: PLoS ONE

    Article Title: Phage-Derived Fully Human Monoclonal Antibody Fragments to Human Vascular Endothelial Growth Factor-C Block Its Interaction with VEGF Receptor-2 and 3

    doi: 10.1371/journal.pone.0011941

    Figure Lengend Snippet: Affinity matured anti-VEGF-C scFvs possess a higher affinity. (A, B) ELISA analysis of bacterial supernatant from randomly picked affinity matured clones after 1 to 3 rounds of selection on biotinylated (A) P. pastoris -derived or (B) mammalian cell-derived ΔNΔC-VEGF-C. (C) BIAcore profiles of monomeric affinity matured anti-VEGF-C scFvs. Monomeric fractions of protein-A purified scFv were prepared by FPLC and injected as 2-fold dilution series on a streptavidin-sensorchip coated with 2000 RU biotinylated ΔNΔC-VEGF-C derived from mammalian cells.

    Article Snippet: Prior to selection, 50 µl streptavidin coated magnetic beads (M-280 Dynabeads, Dynal Biotech, Oslo, Norway) per selection were blocked with 1 ml 3% BSA in PBS for 1 hour at room temperature and resuspended in 50 µl 2% BSA/PBS.

    Techniques: Enzyme-linked Immunosorbent Assay, Clone Assay, Selection, Derivative Assay, Purification, Fast Protein Liquid Chromatography, Injection