Rabbit igg control polyclonal antibody

CoraLite^®488-conjugated CD9 monoclonal antibody (Catalog number: CL488-60232), HRP-conjugated GFP monoclonal antibody (Catalog number: HRP-66002), rabbit IgG control polyclonal antibody (Catalog number: 30000-0-AP), mouse antibody against β-actin (Catalog number: 66009-1-Ig), and rabbit antibodies against VPS25 (Catalog number: 15669-1-AP), GFP (Catalog number: 50430-2-AP), HRS (Catalog number: 10390-1-AP), and Tsg101 (Catalog number: 28283-1-AP) were purchased from ProteinTech Group (Wuhan, China). Mouse antibody against CD63 (Catalog number: sc-5275) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit antibodies against CD9 (Catalog number: 98327S) and calnexin (Catalog number: 2679) were purchased from Cell Signaling Technology (Beverly, MA, USA). Rabbit antibody against HA (Catalog number: H6908) and mouse antibody against FLAG (Catalog number: F3165) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit antibody against EV71 VP1 (Catalog number: GTX132339) was purchased from GeneTex, Inc. (Irvine, CA, USA). Rabbit antibody against EV71 3C (Catalog number: A10003) was purchased from ABclonal Technology (Wuhan, China). Mouse antibody against EV71 3A was kindly provided by Dr. Yongbo Yang of Central China Normal University.

For detecting the interaction between E2 and EGFR, Huh 7 cells were infected with HCV (MOI = 10) for the indicated time. And then cells were gently scraped off the bottom of the cell dish into the PBS using a cell scraper. For cell-surface staining, cell suspensions were washed twice in PBS and stained with the indicated antibodies for 30 min on ice and washed with PBS. For intracellular staining, the cells were fixed for 0.5 h (fixation buffer, BioLegend, Cat# 420801), permeabilized for 10 min (Perm Wash Buffer, BioLegend, Cat# 421002), and then stained with the indicated antibodies. All FCM analysis was conducted on CytoFlex (Beckman), and the data were analyzed using FlowJo V10, according to manufacturers’ instructions. Antibodies used for flow cytometry analysis: HCV-E2 (dilution 1:50, SinoBiological, China, Cat# 40280-T62), EGFR (dilution 1:100, Proteintech, USA, Cat# 66455-1-Ig). Isotype control antibodies (Rabbit IgG control Polyclonal antibody, Proteintech, USA, Cat# 30000-0-AP; Mouse IgG1 isotype control monoclonal antibody, Proteintech, USA, Cat# 66360-1-Ig) were used to define background and non-specific binding signal.

The SMFA was carried out as described previously (49 ). In short, the polyclonal anti-Pfs16 antibody was generated in rabbits and purified with immunogen affinity (Boster Bio). A 15- to 17-day-old P. falciparum culture was used to infect the mosquitoes with purified anti-Pfs16 rabbit polyclonal antibody in 1× PBS mixed with infectious blood (final antibody concentrations are 0.1 mg/ml, 0.2 mg/ml, and 0.4 mg/ml, respectively). The purified rabbit IgG control polyclonal antibody (proteintech) was used as the control to keep total antibody concentration at 0.4 mg/ml. After feeding, only the engorged mosquitoes were kept and maintained on a 10% sucrose solution. Seven days postinfection, midguts were dissected, stained with 0.1% mercurochrome, and examined using a light microscope to count oocysts. In case of the protein feeding assay, mosquitoes were fed with purified rPfs16. The rPfs16 protein was generated and purified as described in this article.