mouse anti human fh mab  (Bio-Rad)

Journal: The Journal of infectious diseases

Article Title: Outer membrane protein OlpA contributes to Moraxella catarrhalis serum resistance via interaction with factor H and the alternative pathway.

doi: 10.1093/infdis/jiu241

Figure Lengend Snippet: Figure 1. Moraxella catarrhalis binds factor H (FH) and OlpA is the main FH-binding protein. A, Most M. catarrhalis strains bind FH, as demonstrated by far Western blotting. B, Identification of 2 FH-binding outer membrane proteins with sizes of approximately 24 kDa and 20 kDa were identified in a 2- dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In panels A, B, and D, whole-cell bacterial lysates or outer membrane vesicles of M. catarrhalis were separated by SDS-PAGE or 2-dimensional SDS-PAGE and either stained by Coomassie blue or transferred to membranes followed by incubation with human complement FH and an anti-FH monoclonal antibody. C and F, M. catarrhalis strains were incubated with FH followed by fluorescein isothiocyanate (FITC)–conjugated anti-FH antibody and flow cytometry. The negative controls (background) represent bacteria incubated with FITC-conjugated antibody only. E, Bacteria were incubated with FH (10 µg/mL) followed by a FITC-conjugated anti-FH antibody, and thereafter bacteria were fixed on glass slides. Representative experiments are shown. In panel F, the mean of 3 values is presented, and error bars represent standard deviations. **P < .01. Abbreviation: wt, wild type.

Article Snippet: Membranes were thereafter incubated with a monoclonal mouse anti-human complement FH antibody (AbD Serotec) diluted 1:1000, followed by horseradish peroxidase–conjugated rabbit anti-mouse polyclonal antibodies (Dako).

Techniques: Binding Assay, Far Western Blot, Membrane, Polyacrylamide Gel Electrophoresis, SDS Page, Staining, Incubation, Cytometry, Bacteria

Journal: PLoS ONE

Article Title: Comparative Proteomic Analysis of Serum from Patients with Systemic Sclerosis and Sclerodermatous GVHD. Evidence of Defective Function of Factor H

doi: 10.1371/journal.pone.0012162

Figure Lengend Snippet: ( A ) Immuno-blot analysis of PBP and graph reporting the quantification of PBP derived protein expression as detected by densitometric analysis. 35 µg of total serum proteins were solubilised under reducing conditions, loaded on 12% polyacrylamide gels, transferred to nitrocellulose membranes and probed with goat polyclonal anti-PBP antibody. We could not separate by this technique the individual peptides of this family because of their small differences in the amino acid sequence length. Shown is one representative experiment of five with similar results. ( B ) Immuno-blot analysis of FH and graph reporting the quantification of FH expression as detected by densitometric analysis. FH molecules from serum of dSSc, lSSc, ScGVHD patients and H, bT and T w/o GVHD subjects were separated by mono-dimensional electrophoresis on 8% polyacrylamide gels under non reducing conditions. Samples were transferred to nitrocellulose membranes and probed with mouse monoclonal anti-FH antibody. Shown is one representative experiment of five with similar results.

Article Snippet: Cells were treated with patient and control sera (20% in 0.5xDPBS) for 20 min at 4°C and subsequently incubated with mouse monoclonal anti- human FH antibody (AbD Serotec) and Alexa-fluor 488–conjugated goat anti-mouse IgG (Invitrogen) 1∶100 at 4°C for 20 min. All washes and incubation steps were performed in 0.5xPBS containing 0.5% BSA.

Techniques: Derivative Assay, Expressing, Sequencing, Electrophoresis