detection antibody  (Cedarlane)

Journal: Applied microbiology and biotechnology

Article Title: A new standardization for the use of chicken embryo: selection of target from the phage display library and infection.

doi: 10.1007/s00253-024-13227-x

Figure Lengend Snippet: Fig. 1 Percentage of embryos killed with different doses of SP at two different ages. NC, negative control group with PBS; 6 log/embryo, group inoculated with 6 log/embryo of SP; 4 log/embryo, group inoc- ulated with 4 log/embryo of SP; 2 log/embryo, group inoculated with 2 log/embryo of SP

Article Snippet: Then, we incubate the plates with the antibodies, rabbit anti-chicken IFN-γ IgG antibody (BioRad), rabbit anti-chicken IL-1β IgG antibody (BioRad) or IL-10 Polyclonal IgG antibody (Thermo), diluted (1:500) in 3% BSA + PBS for 1 h at 37 °C.

Techniques: Negative Control

Journal: BMC microbiology

Article Title: Single stranded (ss)RNA-mediated antiviral response against infectious laryngotracheitis virus infection.

doi: 10.1186/s12866-019-1398-6

Figure Lengend Snippet: Fig. 1 In ovo treated resiquimod increases macrophage recruitment in tissues of respiratory and gastrointestinal systems. SPF eggs were treated in ovo with resiquimod (n = 5) or PBS (n = 4) at ED18 and the eggs were incubated until hatch. At day 1 post-hatch, trachea, lungs, duodenum and large intestine were preserved in OCT, sectioned and immunofluorescent assay was performed with mouse monoclonal antibody specific for chicken macrophages, KUL01 (red signals) and with DAPI (blue signals) for nuclear staining. The quantitative data and representative immunofluorescent figures from each organ is shown, a lungs, b tracheas, c duodenum and d large intestine. Student’s t-test was performed to identify group differences and the differences were considered significant at P < 0.05

Article Snippet: Then, unlabeled rabbit polyclonal antibody specific for chicken IL-1β (Bio-Rad Laboratories, Mississauga ON, Canada) were used in, 1:10 dilutions in block buffer and incubated for 30 min. As a secondary antibody for IL-1β, biotinylated goat anti-rabbit IgG(H + L) (Vectashield, Vector Laboratories Inc., Burlingame, CA, USA) was used in 1:400 dilution in block buffer and incubated for 20 min and subsequently incubated with DyLight® 488 for 20 min. TBS-T buffer (TBS with 0.1% Tween 20) was used to wash the slides twice, followed by a single wash with PBS following each step.

Techniques: In Ovo, Incubation, Staining

Journal: BMC microbiology

Article Title: Single stranded (ss)RNA-mediated antiviral response against infectious laryngotracheitis virus infection.

doi: 10.1186/s12866-019-1398-6

Figure Lengend Snippet: Fig. 5 Stimulation of avian macrophages with the synthetic ssRNA, resiquimod increases the production of IL-1β but not type 1 IFNs. a Avian macrophages, MQ-NCSU cells, were cultured in 12-well plates (1 × 106 cells per well) for 24 h and stimulated with either resiquimod (10 μg/ml) or growth media (control) in triplicates. The culture supernatants were collected 24 h post-treatment and transferred to DF-1 cell monolayers grown in 12-well plates and infected with VSV conjugated with GFP (0.1 MOI). At 24 h post infection, the percentage of cells expressing GFP signal was quantified after fixation with 4% paraformaldehyde and nuclear staining. The experiment was repeated two more times with similar results and the data were pooled. The representative figures in each group are shown. b The MQ-NCSU cells were cultured on coverslips in 12-well plates with 1 × 106 viable cells per well and protein transport inhibitor (2 μl/ml) was added to culture medium after 6 h. After 24 h of culture, the cells were stimulated with resiquimod (10 μg/ml), or RPMI media as a control separately (triplicates per treatment). Following 24 h of treatment, immunofluorescent staining targeting IL-1β was performed after fixation with 4% paraformaldehyde. The experiment was repeated two more times with similar results and the data were pooled. The representative figures in each group are shown. Student’s t-test was used to identify differences between two groups and the differences were considered significant at P < 0.05

Article Snippet: Then, unlabeled rabbit polyclonal antibody specific for chicken IL-1β (Bio-Rad Laboratories, Mississauga ON, Canada) were used in, 1:10 dilutions in block buffer and incubated for 30 min. As a secondary antibody for IL-1β, biotinylated goat anti-rabbit IgG(H + L) (Vectashield, Vector Laboratories Inc., Burlingame, CA, USA) was used in 1:400 dilution in block buffer and incubated for 20 min and subsequently incubated with DyLight® 488 for 20 min. TBS-T buffer (TBS with 0.1% Tween 20) was used to wash the slides twice, followed by a single wash with PBS following each step.

Techniques: Cell Culture, Control, Infection, Expressing, Staining

Journal: BMC microbiology

Article Title: Single stranded (ss)RNA-mediated antiviral response against infectious laryngotracheitis virus infection.

doi: 10.1186/s12866-019-1398-6

Figure Lengend Snippet: Fig. 6 Antiviral activity of synthetic resiquimod against ILTV replication is not dependent on IL-1β production. Avian macrophages, MQ-NCSU cells, were cultured in 12-well plates (1 × 106 cells per well) for 24 h and stimulated with either resiquimod (10 μg/ml) or growth media (control) in 6 replicates. The resultant MQ-NCSU cell culture supernatants were collected at 24 h post-treatment and transferred (250 μl) to LMH cell monolayer. Before transferring MQ-NCSU cell culture supernatants, 3 wells from resiquimod and control groups in LMH cells were incubated with 1.0 μg/ml of IL-1Ra for 30 min. Twenty-four hours later, the LMH cells were infected with ILTV (300 PFU/well). The plates were stained with 1% crystal violet at 5 dpi and resulting plaques were counted. The experiment was repeated two more times with the same number of replicates with similar results and the data were pooled. ANOVA with Bonferroni’s posttest for selected comparison was performed to identify the differences between groups and the differences were considered significant at P < 0.05

Article Snippet: Then, unlabeled rabbit polyclonal antibody specific for chicken IL-1β (Bio-Rad Laboratories, Mississauga ON, Canada) were used in, 1:10 dilutions in block buffer and incubated for 30 min. As a secondary antibody for IL-1β, biotinylated goat anti-rabbit IgG(H + L) (Vectashield, Vector Laboratories Inc., Burlingame, CA, USA) was used in 1:400 dilution in block buffer and incubated for 20 min and subsequently incubated with DyLight® 488 for 20 min. TBS-T buffer (TBS with 0.1% Tween 20) was used to wash the slides twice, followed by a single wash with PBS following each step.

Techniques: Activity Assay, Cell Culture, Control, Transferring, Incubation, Infection, Staining, Comparison

Journal: BMC microbiology

Article Title: Single stranded (ss)RNA-mediated antiviral response against infectious laryngotracheitis virus infection.

doi: 10.1186/s12866-019-1398-6

Figure Lengend Snippet: Fig. 7 In ovo synthetic resiquimod treatment increases the production of IL-1β from macrophages in lungs in vivo. SPF eggs were treated in ovo with resiquimod (n = 5) or PBS (n = 4) at ED18 and the eggs were incubated until hatch. At day 1 post-hatch, lungs were preserved in OCT, sectioned and double immunofluorescent assay was performed to identify the macrophages that produce IL-1β. The quantitative data from lung is shown along with representative immunoassayed images. Student’s t-test was performed to identify group differences and the differences were considered significant at P < 0.05

Article Snippet: Then, unlabeled rabbit polyclonal antibody specific for chicken IL-1β (Bio-Rad Laboratories, Mississauga ON, Canada) were used in, 1:10 dilutions in block buffer and incubated for 30 min. As a secondary antibody for IL-1β, biotinylated goat anti-rabbit IgG(H + L) (Vectashield, Vector Laboratories Inc., Burlingame, CA, USA) was used in 1:400 dilution in block buffer and incubated for 20 min and subsequently incubated with DyLight® 488 for 20 min. TBS-T buffer (TBS with 0.1% Tween 20) was used to wash the slides twice, followed by a single wash with PBS following each step.

Techniques: In Ovo, In Vivo, Incubation