rabbit polyclonal ab  (GE Healthcare)

Journal: PLoS Pathogens

Article Title: Immunogenetic Mechanisms Driving Norovirus GII.4 Antigenic Variation

doi: 10.1371/journal.ppat.1002705

Figure Lengend Snippet: Predicted GII.4 norovirus evolving blockade epitopes. Bioinformatic approaches predicted five antibody epitopes on the surface of GII.4 noroviruses that appeared to be evolving over time and to correlate with the emergence of new GII.4 outbreak strains. Panel A : Amino acid variation of Epitopes A–E by GII.4 strain. Panel B : Predicted epitopes were expanded to include interacting amino acids within 8A. Epitope A (grey), Epitope B (blue), Epitope C (green), Epitope D (black), Epitope E (teal) and HBGA binding sites (magenta) mapped onto the P domain dimer of GII.4.2002.

Article Snippet: Bound VLP was detected by a rabbit anti-GII norovirus polyclonal sera made from hyperimmunization with either GII.4.2009 or a cocktail of GII.4.1997, GII.3.1999, GII.1.1976, and GII.2.1976 VLPs, followed by anti-rabbit IgG-HRP (GE Healthcare) and color developed with 1-Step Ultra TMB ELISA HRP substrate solution (Thermo-Fisher).

Techniques: Binding Assay

Journal: PLoS Pathogens

Article Title: Immunogenetic Mechanisms Driving Norovirus GII.4 Antigenic Variation

doi: 10.1371/journal.ppat.1002705

Figure Lengend Snippet: Characterization of donor NVB plasma blockade of norovirus VLPs. Panel A : PGM binding blockade activity. Sigmoidal curves were fit to the mean percent control binding calculated by comparing the amount of VLP bound to PGM in the presence of antibody pretreatment to the amount of VLP bound in the absence of antibody pretreatment. Error bars represent the standard error of the mean. Panel B : Mean EC50 (% plasma) for blockade of each VLP. * VLPs with EC50 values significantly different from the EC50 for GII.4.2006.

Article Snippet: Bound VLP was detected by a rabbit anti-GII norovirus polyclonal sera made from hyperimmunization with either GII.4.2009 or a cocktail of GII.4.1997, GII.3.1999, GII.1.1976, and GII.2.1976 VLPs, followed by anti-rabbit IgG-HRP (GE Healthcare) and color developed with 1-Step Ultra TMB ELISA HRP substrate solution (Thermo-Fisher).

Techniques: Binding Assay, Activity Assay

Journal: Journal of Virology

Article Title: Emergence of a Norovirus GII.4 Strain Correlates with Changes in Evolving Blockade Epitopes

doi: 10.1128/JVI.03106-12

Figure Lengend Snippet: Epitope A constitutes ∼40% of the polyclonal blockade antibody response in NoV outbreak human convalescent-phase serum. Convalescent serum samples collected from eight GII.4.2009-infected subjects were assayed for ability to block VLP interaction

Article Snippet: VLPs (0.5 μg/ml) were pretreated with decreasing concentrations of test MAb or outbreak sera for 1 h at room temperature before being added to the carbohydrate ligand-coated plates for 1 h. Bound VLP were detected by a rabbit anti-GII norovirus polyclonal sera made from hyperimmunization with either GII.4.2009 or a cocktail of GII.4.1997, GII.3.1999, GII.1.1976, and GII.2.1976 VLPs, followed by anti-rabbit IgG-HRP (GE Healthcare) and color developed with 1-Step Ultra TMB ELISA HRP substrate solution (Thermo Fisher).

Techniques: Infection, Blocking Assay

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: N-terminal proteolytic processing of PAP-III in injured sciatic nerves of rats. A , induction of PAP-III mRNA in injured sciatic nerves. Nerves were collected 0 ( cont ), 1, 3, and 7 days ( d ) after axotomy. Expression of PAP-III mRNA was examined by RT-PCR.

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: Weak interactions of exogenously added ΔN-PAP-III fibers with glial cells. ΔN-PAP-III fibers produced in 25 m m Tris (pH 7.5) containing 150 m m NaCl were added to the culture media of astrocytes, microglia, and Schwann cells, and incubated

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques: Produced, Incubation

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: Association of exogenously added ΔN-PAP-III fibers with neurons. ΔN-PAP-III fibers produced in 25 m m Tris (pH 7.5) containing 150 m m NaCl were added to the culture media of primary cortical neurons precultured for 12–18 h. Twenty-four

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques: Produced

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: Precipitability of ΔN-PAP-III depends on Na + concentrations. A , PAP-III protein was incubated with (+) or without (−) trypsin in 25 m m Tris (pH 7.5) containing 0 (−), 25, and 150 m m NaCl for 2 h ( hrs ). B , PAP-III protein was treated

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques: Incubation

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: No significant effects of exogenously added full-length PAP-III or PAP-III (N) on neurite extension. Primary cortical neurons were seeded on PDL-coated glass coverslips and cultured for 36 h in media containing no PAP-III ( control ), full-length PAP-III,

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques: Cell Culture

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: Neurite extension on ΔN-PAP-III fibers immobilized on glass coverslips. Dense matrices of ΔN-PAP-III fibers were prepared on PDL-coated glass coverslips by digesting ΔN-PAP-III without NaCl. Primary cortical neurons were seeded

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: N-terminal Cleaved Pancreatitis-associated Protein-III (PAP-III) Serves as a Scaffold for Neurites and Promotes Neurite Outgrowth *

doi: 10.1074/jbc.M112.395301

Figure Lengend Snippet: Fibrillar structure of ΔN-PAP-III. A , PAP-III treated with trypsin in 25 m m Tris (pH 7.5) containing 0 (−), 25, and 150 m m NaCl was attached to PDL-coated coverslips and visualized by immunofluorescence using anti-PAP-III antibody. Scale

Article Snippet: The membrane was probed with a mouse monoclonal anti-GAPDH antibody (4300; Ambion, Huntingdon, UK) or a rabbit polyclonal anti-PAP-III antibody produced in our previous study and then with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) ( ).

Techniques: Immunofluorescence

Journal: Molecular cell

Article Title: Multiple Functions for Mcm2-7 ATPase Motifs During Replication Initiation

doi: 10.1016/j.molcel.2014.06.033

Figure Lengend Snippet: Mcm2-7 ATPase motif mutants are defective for in vitro DNA replication. Replication assays were performed with purified wild-type or Mcm2-7 ATPase motif mutant complexes. Radiolabeled DNA replication products were analyzed by alkaline agarose electrophoresis

Article Snippet: Eluted proteins were incubated with 30 µl anti-Mcm2-7 Ab (UM174, rabbit polyclonal) bound to Gammabind G Sepharose (GE) in 400 µl Buffer H containing 0.3 M KGlu for 90 minutes.

Techniques: In Vitro, Purification, Mutagenesis, Electrophoresis

Journal: Molecular cell

Article Title: Multiple Functions for Mcm2-7 ATPase Motifs During Replication Initiation

doi: 10.1016/j.molcel.2014.06.033

Figure Lengend Snippet: Initial association of ATPase motif mutant Mcm2-7/Cdt1 complexes. Helicaseloading assays were performed in the presence of ATPγS followed by a low-salt wash allowing detection of the initial DNA association of Mcm2-7 and helicase loading proteins

Article Snippet: Eluted proteins were incubated with 30 µl anti-Mcm2-7 Ab (UM174, rabbit polyclonal) bound to Gammabind G Sepharose (GE) in 400 µl Buffer H containing 0.3 M KGlu for 90 minutes.

Techniques: Mutagenesis

Journal: Molecular cell

Article Title: Multiple Functions for Mcm2-7 ATPase Motifs During Replication Initiation

doi: 10.1016/j.molcel.2014.06.033

Figure Lengend Snippet: Loaded Mcm2-7 ATPase motif mutants form double hexamers that are DDK substrates. (A) Double hexamer formation by ATPase motif mutant Mcm2-7 complexes. After a helicase-loading reaction and high-salt wash, loaded wild-type or mutant Mcm2-7 complexes were

Article Snippet: Eluted proteins were incubated with 30 µl anti-Mcm2-7 Ab (UM174, rabbit polyclonal) bound to Gammabind G Sepharose (GE) in 400 µl Buffer H containing 0.3 M KGlu for 90 minutes.

Techniques: Mutagenesis

Journal: Molecular cell

Article Title: Multiple Functions for Mcm2-7 ATPase Motifs During Replication Initiation

doi: 10.1016/j.molcel.2014.06.033

Figure Lengend Snippet: Mcm2-7 ATPase motif mutants exhibit defects in multiple steps during helicase activation. (A) Mcm2-7 ATPase mutants are defective in DNA retention during helicase activation. The amount of the indicated Mcm2-7 complexes (Flag-Mcm3) retained on DNA was

Article Snippet: Eluted proteins were incubated with 30 µl anti-Mcm2-7 Ab (UM174, rabbit polyclonal) bound to Gammabind G Sepharose (GE) in 400 µl Buffer H containing 0.3 M KGlu for 90 minutes.

Techniques: Activation Assay

Journal: Molecular cell

Article Title: Multiple Functions for Mcm2-7 ATPase Motifs During Replication Initiation

doi: 10.1016/j.molcel.2014.06.033

Figure Lengend Snippet: Mcm2-7 ATPase motif mutants are defective in helicase loading. (A) Helicase loading of MCM ATPase motif mutants was monitored by incubating loading proteins with ATP and origin-DNA-beads followed by a high-salt wash. The associated graph shows the relative

Article Snippet: Eluted proteins were incubated with 30 µl anti-Mcm2-7 Ab (UM174, rabbit polyclonal) bound to Gammabind G Sepharose (GE) in 400 µl Buffer H containing 0.3 M KGlu for 90 minutes.

Techniques: