human recombinant c5a desarg  (Hycult Biotech)

Journal: Oncotarget

Article Title: A novel multiplex detection array revealed systemic complement activation in oral squamous cell carcinoma

doi: 10.18632/oncotarget.22963

Figure Lengend Snippet: ( A ) Serial dilutions of native, human C3a-desArg (black circle), recombinant, human C5a-desArg (black square) and native human sC5b-C9 (black triangle) depicted different assay performances for the simultaneous analysis of complement activation markers. The target antigens were only detected by respective coupled magnetic beads and not by control beads (grey). Limit of detection (orange) and assay cutoffs (red square) for each analyte are shown. ( B ) The concentration range of complement activation markers determined with the novel multiplex assay from a human serum pool (black square with dot) corresponded to previously reported ranges for C3a, C5a and sC5b-C9 single detection either from plasma (grey), serum (black) or unknown sample matrix (white), respectively [ – ].

Article Snippet: A mixture of serial 1:3 diluted human native C3a-desArg (197.5–0.001 ng/mL, cat. HC2127, Hycult Biotech, Beutelsbach, Germany), human recombinant C5a-desArg (28.1–0.004 ng/mL, Hycult Biotech, Beutelsbach, Germany) and human native sC5b-9 (66.6–0.0004 µg/mL, cat. A127, Complement Technology, Tyler, TX, USA) were used as standard curve.

Techniques: Recombinant, Activation Assay, Magnetic Beads, Concentration Assay, Multiplex Assay

Journal: Journal of Immunology Research

Article Title: Venom from Bothrops lanceolatus , a Snake Species Native to Martinique, Potently Activates the Complement System

doi: 10.1155/2018/3462136

Figure Lengend Snippet: Production of Complement activation products in NHS incubated with B. lanceolatus venom. NHS (diluted 1 : 2 in saline solution) was incubated with B. lanceolatus venom (0.5 mg/mL) for 30 min at 37°C, in the presence or absence of 1,10-phenantroline (10 mM). As control, NHS was incubated with saline solution. (a) Samples were diluted 1 : 5000 and the concentrations of the anaphylatoxins C3a/C3a-desArg, C4a/C4a-desArg, and C5a/C5a-desArg were determined using a cytometric bead array. (b) Samples were diluted 1 : 150, and the concentration of the SC5b-9 complex was measured by “MicroVue SC5b-9 Plus EIA Kit”. The data represent two experiments, realized in duplicate. ∗ p < 0.05 compared to the saline-treated NHS controls. # p < 0.05 compared to venom-treated samples.

Article Snippet: C3a/C3a-desArg, C4a/C4a-desArg, and C5a/C5a-desArg were measured using the Human Anaphylatoxin Cytometric Bead Array (CBA) (BD Biosciences Pharmingen, San Diego, CA, USA), following the manufacturer's instructions.

Techniques: Activation Assay, Incubation, Concentration Assay

Journal: Oncotarget

Article Title: A novel multiplex detection array revealed systemic complement activation in oral squamous cell carcinoma

doi: 10.18632/oncotarget.22963

Figure Lengend Snippet: ( A ) Serial dilutions of native, human C3a-desArg (black circle), recombinant, human C5a-desArg (black square) and native human sC5b-C9 (black triangle) depicted different assay performances for the simultaneous analysis of complement activation markers. The target antigens were only detected by respective coupled magnetic beads and not by control beads (grey). Limit of detection (orange) and assay cutoffs (red square) for each analyte are shown. ( B ) The concentration range of complement activation markers determined with the novel multiplex assay from a human serum pool (black square with dot) corresponded to previously reported ranges for C3a, C5a and sC5b-C9 single detection either from plasma (grey), serum (black) or unknown sample matrix (white), respectively [ – ].

Article Snippet: Purified recombinant human C3a-desArg (1 µg, HC2127), C5a-desArg (1 µg, HC2102, Hycult Biotech, Beutelsbach, Germany), sC5b-9 (1 µg/0.1 µg, A127, Complement Technology, Tyler, TX, USA) or normal human serum (20 µg, NHS) were separated under reducing and non-reducing conditions in either 10% or 15% SDS-PAGE and subsequently transferred to polyvinylidene difluoride membranes.

Techniques: Recombinant, Activation Assay, Magnetic Beads, Concentration Assay, Multiplex Assay