Journal: Cell reports

Article Title: Mining HIV controllers for broad and functional antibodies to recognize and eliminate HIV-infected cells

doi: 10.1016/j.celrep.2021.109167

Figure Lengend Snippet:

Article Snippet: Mouse Anti-Human C3 / C3b / iC3b Monoclonal Antibody, FITC Conjugated , CedarLane , CEDARLANE Cat# CL7632F; RRID:AB_10548984.

Techniques: Control, Virus, Recombinant, Reporter Gene Assay, Staining, Labeling, Software

Journal: International Journal of Nanomedicine

Article Title: Comparative in vitro and in vivo Evaluation of Different Iron Oxide-Based Contrast Agents to Promote Clinical Translation in Compliance with Patient Safety

doi: 10.2147/IJN.S402320

Figure Lengend Snippet: Contrast agent-induced complement activation. ( A ) Determination of the iC3b split product, in human-derived platelet-poor plasma (n=3). Relative hemodynamic changes as indication of in vivo complement activation in a porcine model after administration of SPION Dex ( B ), ferucarbotran ( C ), and ferumoxytol ( D ) at an iron concentration of 5 mg/kg. The latter resulted in anaphylaxis and requiring subsequent cardiopulmonary resuscitation. ( E ) Comparisons of maximum pulmonary artery pressure after contrast agent application (data for both doses, 0.5 and 5.0 mg Fe/kg, are pooled). ( F ) Normalized thromboxane B2 levels in pigs acquired from blood samples collected during application. ( G ) Example image of flushes appearing in the abdominal region after ferumoxytol application.

Article Snippet: Complement activation: To analyze complement activation, an iC3b ELISA obtained from MicroVue (Quidel, USA) was performed.

Techniques: Activation Assay, Derivative Assay, Clinical Proteomics, In Vivo, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Functional Analysis of Variants in Complement Factor I Identified in Age-Related Macular Degeneration and Atypical Hemolytic Uremic Syndrome

doi: 10.3389/fimmu.2021.789897

Figure Lengend Snippet: Systemic complement levels in CFI rare variant carriers. (A) FI, (B) FH, and (C) C3bBbP plasma concentrations and (D) the amount of iC3b generated in the C3b degradation assay were determined with ELISA. The C3b degradation was performed three times, and iC3b levels were determined in duplicate. Dotted lines indicate the mean concentrations ± 2SD determined in non-carriers. The mean ± 2SD is (A) 57 µg/ml ± 11 µg/ml for FI, (B) 390 µg/ml ± 64 µg/ml for FH, (C) 12.9 CAU/ml ± 4.1 CAU/ml for C3bBbP, and (D) 8.5 µg/ml ± 1.6 µg/ml for generated iC3b. Solid lines indicate the mean concentration for each group.

Article Snippet: Then, the samples were immediately diluted 40x in PBS with 0.02% Tween and applied to high binding microplates (Greiner, Austria) coated with mouse-anti iC3b (A209, Quidel, USA) 2,500x diluted in coating buffer (15 mM Na 2 CO 3 *10 H 2 O, 35 mM NaHCO 3 , pH 9.6) and blocked with 1% BSA (Sigma, USA).

Techniques: Variant Assay, Clinical Proteomics, Generated, Degradation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Functional Analysis of Variants in Complement Factor I Identified in Age-Related Macular Degeneration and Atypical Hemolytic Uremic Syndrome

doi: 10.3389/fimmu.2021.789897

Figure Lengend Snippet: CFI variants affect C3b degradation. C3b degradation was performed and generated iC3b was measured with ELISA. 15 µg/ml C3b and 3 µg/ml FH were added to purified FI, and incubated for 3h at 37°C. Th negative control does not contain purified FI, and for the positive control 10 ug/ml serum purified FI was incubated with the C3b/FH mix. As additional control, recombinant WT FI was incubated with 15 µg/ml C3, but FH omitted (WT -FH). The amount of iC3b generated by the variants was normalized to the concentration generated by recombinantly expressed WT FI. Bars and error bars show the mean and SD, individual dots represent the individual measurements. Samples were compared to WT using one-way ANOVA followed by Dunnet’s multiple comparison test. P-values are indicated as ****<0.001.

Article Snippet: Then, the samples were immediately diluted 40x in PBS with 0.02% Tween and applied to high binding microplates (Greiner, Austria) coated with mouse-anti iC3b (A209, Quidel, USA) 2,500x diluted in coating buffer (15 mM Na 2 CO 3 *10 H 2 O, 35 mM NaHCO 3 , pH 9.6) and blocked with 1% BSA (Sigma, USA).

Techniques: Generated, Enzyme-linked Immunosorbent Assay, Purification, Incubation, Negative Control, Positive Control, Control, Recombinant, Concentration Assay, Comparison