Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Therapeutic targeting of vasculature in the premetastatic and metastatic niches reduces lung metastasis

doi: 10.4049/jimmunol.1901208

Figure Lengend Snippet: (A) C5a concentration in bronchoalvolar lavage from tumor-free (TF) and mice with 4T1 tumors at day 8, *P<0.05 by t-test; Immunofluorescent detection of (B) C3 cleavage fragments and CD31, (C) C3 cleavage fragments and C1q-arrow indicates colocalization, (D) C1q and IgM, and (E) IgM and Annexin V in the lungs of tumor bearing mice at the day 8 (arrows indicate colocalization). Scale bars 50 μm. Data are representative of one experiment with at least 4 mice per cohort.

Article Snippet: Immunofluorescence Frozen tissue sections 5-μm thick were stained with: CD31 (Clone 390, BD Pharmingen), CD8a (53–6.7, BD Pharmingen), NG2 (546930, R&D Systems), CD105 (MJ7/18, Biolegend), Ki-67 Ki-67 (D3B5, Cell Signaling), C3b/iC3b/C3c (HM 1065, Hycult Biotech), C1q (HM 1096BT, Hycult), mannose-binding lectin (NB100–1502 Novus), IgM (14–5790-81, eBioscience), and IgG (406705, Biolegend) antibodies.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: The Prolonged Treatment of Salmonella enterica Strains with Human Serum Effects in Phenotype Related to Virulence

doi: 10.3390/ijms24010883

Figure Lengend Snippet: Analysis of complement C1q ( A ) and C3c ( B ) deposition on S . enterica cells. Cells of the WT serum-sensitive (SS) strains and the serum-resistant (SR, P8) were incubated in 50% SPPP for 12 min at 37 °C. C1q and C3c components deposited on cells were identified by indirect ELISA with appropriate antibodies as described in the methods. Data are presented as the means of standard deviations of triplicate determinations from three independent experiments, after subtraction of negative control (A 490 0.125); * indicates a statistically significant difference in values ( p < 0.05) with respect to the data obtained for WT strains.

Article Snippet: The plates were incubated with mouse anti-human C1q monoclonal antibody (1:1000 dissolved in 1% low-fat milk) (GeneTex, Irvine, CA, USA, 1.0 mg/mL −1 ), then with donkey anti-mouse IgG antibody (1:1000 dissolved in 1% low-fat milk)/HRP (horseradish peroxidase-conjugated, GeneTex, 1.0 mg/mL −1 ) and finally with o -phenylenediamine dihydrochloride (Thermo Fisher Scientific, Waltham, MA, USA) to develop alkaline phosphatase and measured at 490 nm.

Techniques: Incubation, Indirect ELISA, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: The Prolonged Treatment of Salmonella enterica Strains with Human Serum Effects in Phenotype Related to Virulence

doi: 10.3390/ijms24010883

Figure Lengend Snippet: Dot blot C1q and C3c-binding assays performed for membrane proteins (MPs) isolated from serum-resistant (SR) and serum-sensitive (SS) strains. 1—MPs of SS S . Hammonia, 2—MPs of SR S . Hammonia P8, 3—MPs of SS S . Typhimurium ATCC 14028, 4—MPs of SS S . Typhimurium ATCC 14028 P8. MPs in concentrations of ( A ) (25 × diluted in H 2 O miliQ , 0.24 mg/mL −1 ) and ( B ) (50 × diluted in H 2 O miliQ, 0.12 mg/mL −1 ) were loaded onto nitrocellulose membrane. K1—positive control 1; K2—positive control 2; K3—negative control 1, instead of MPs PBS was used; K4, negative control 2—instead of SPPP PBS was used.

Article Snippet: The plates were incubated with mouse anti-human C1q monoclonal antibody (1:1000 dissolved in 1% low-fat milk) (GeneTex, Irvine, CA, USA, 1.0 mg/mL −1 ), then with donkey anti-mouse IgG antibody (1:1000 dissolved in 1% low-fat milk)/HRP (horseradish peroxidase-conjugated, GeneTex, 1.0 mg/mL −1 ) and finally with o -phenylenediamine dihydrochloride (Thermo Fisher Scientific, Waltham, MA, USA) to develop alkaline phosphatase and measured at 490 nm.

Techniques: Dot Blot, Binding Assay, Membrane, Isolation, Positive Control, Negative Control