mouse anti human ig λ  (SouthernBiotech)

Journal: PLOS Biology

Article Title: A flexible loop in the paxillin LIM3 domain mediates its direct binding to integrin β subunits

doi: 10.1371/journal.pbio.3002757

Figure Lengend Snippet: ( A ) Flow cytometric analysis of NIH Flp-In integrin β3 KO and the indicated ITGB3 wt, ITGB3 Δ8aa, or ITGB3 Δ3aa reexpressing cell lines. Cells were left unstained or stained against mouse integrin β3 or with an isotype matched control IgG. ( B ) WCLs of cell lines in (A) were probed with the indicated antibodies directed against core FA proteins. Tubulin was used as loading control. ( C ) Serum starved cells as in (A) were seeded onto vitronectin-coated (5 μg/ml) glass slides for 30 min, and cell area was measured. Shown are mean values and 95% confidence intervals of n = 60 cells per sample from 3 independent experiments. Statistical significance was calculated using one-way ANOVA followed by Bonferroni multiple comparison test (*** p ≤ 0.001). The data underlying this panel can be found in . ( D ) Flow cytometric analysis of Kindlin 1/2 flox cells (Kind Ctrl ), Kindlin 1/2 KO (Kind KO ), or Kind KO cells stably transduced with either murine full-length integrin β3, truncated integrin β3 Δ8aa or Δ3aa, or empty vector backbone (mock). Cells were analysed for their surface expression of various integrin subunits using flow cytometry. ( E ) WCL from cell lines in (D) were analysed by western blotting with antibodies against indicated core FA proteins. Monoclonal α-tubulin antibody was used as loading control. ( F ) Serum starved cells as in (D) were seeded on glass coverslips coated with 50 μg/ml vitronectin or poly-Lysine for 4 h. Cells were fixed and stained for endogenous paxillin or talin as indicated. Paxillin- and talin-positive cell attachment sites in ITGB3 wt expressing kindlin1/2 KO cells are indicated with black arrowheads.

Article Snippet: The following primary and secondary antibodies were used at indicated concentrations: anti-human α-actinin1 (mouse monoclonal, BM75.2, Sigma Aldrich, A5044; WB 1:1,000), anti-human talin (mouse monoclonal, 8d4, Sigma Aldrich, T3287; WB 1:750), anti-human FAK (rabbit polyclonal, A-17, Santa Cruz, sc-557; WB 1:250), anti-human kindlin2 (mouse monoclonal, 3A3, Merck, MAB2617; WB 1:1,000, IF 1:200), anti-mouse kindlin2 (rabbit polyclonal, 11453-1-AP, Proteintech; WB 1:2,000), anti-human cSRC (rabbit polyclonal, SRC2, Santa Cruz, sc-18; WB 1:1,000), anti-human ILK (rabbit monoclonal, EP1593Y, Epitomics; WB 1:1,000), anti-human p130Cas (rabbit polyclonal, N17, Santa Cruz; WB 1:1,000), anti-human vinculin (mouse monoclonal, VIN-1, Sigma Aldrich, V9131; WB 1:1,000), anti-human Hic-5 (mouse monoclonal, 34, BD Biosciences, 611164; WB 1:500), anti-human paxillin (mouse monoclonal, 5H11, Thermo Fisher Scientific, AHO0492; WB 1:1,000, IF 1:200), anti-GAPDH (mouse monoclonal, GA1R, Thermo Fisher Scientific, MA5-15738-HRP), anti-human Rac (rabbit polyclonal, invitrogen, PA5-17519; WB 1:1,000), anti-human CEACAM1, 3, 4, 5, 6 (mouse monoclonal, D14HD11, Aldevron; WB 1:6,000, IF 1:200); mouse monoclonal anti 6xHis (mouse monoclonal, HIS.H8, Thermo Fisher Scientific, MA1-21315; WB 1:2,000), anti GFP (mouse monoclonal, JL8, Clontech; WB: 1:6,000), anti-human tubulin (mouse monoclonal, E7, purified from hybridoma cell supernatants, Developmental Studies Hybridoma Bank, University of Iowa, USA; WB 1:1,000), anti-mouse integrin β1 (Armenian hamster monoclonal, Hmb1-1, Thermo Fisher Scientific, 11-0291-82; FC 1:300), anti-mouse integrin β3 (Armenian hamster monoclonal, 2C9.G3, Thermo Fisher Scientific, 13-0611-81; FC 1:200), anti-mouse integrin α5 (rat monoclonal, MFR5, BD Biosciences, 553319; FC 1:300), anti-mouse integrin αV (rat monoclonal, RMV-7, BD Biosciences, 550024; FC 1:300).

Techniques: Staining, Control, Comparison, Stable Transfection, Transduction, Plasmid Preparation, Expressing, Flow Cytometry, Western Blot, Cell Attachment Assay

Journal: Cell reports

Article Title: Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38

doi: 10.1016/j.celrep.2024.114502

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Yeast displaying IgGs on their surface were incubated with biotinylated SCP and SMP preps at a 1:10 dilution in PBSF (PBS with 0.1% w/v BSA) and incubated on ice for 20 min. Yeast cells were then washed two times in PBSF and further stained with a cocktail of ExtraAvidin-R-PE (Sigma Aldrich, dilution 1:50), anti-human kappa FITC (Southern Biotech, dilution 1:100), anti-human lambda FITC (Southern Biotech, dilution 1:100), and PI (Invitrogen, dilution 1:100) for 20 min on ice.

Techniques: Derivative Assay, Virus, Recombinant, Protease Inhibitor, Reverse Transcription, Membrane, Cell Isolation, Mutagenesis, Expressing, Plasmid Preparation, Software, Affinity Column, Sequencing, Transfection, Electron Microscopy