human il 13 rα2 biotinylated affinity purified polyclonal detection antibody  (R&D Systems)

Journal: PLOS Pathogens

Article Title: Choline metabolism underpins macrophage IL-4 polarization and RELMα up-regulation in helminth infection

doi: 10.1371/journal.ppat.1011658

Figure Lengend Snippet: A-D) Macrophages were treated with vehicle (DMSO) or HC3 (250 μM) for 24 h, washed, then treated with IL-4 (20 ng/mL) for 24 h. Relative expression of M[IL-4] hallmark genes Arg1 , Mrc1 , Chil3 , or Retnla normalized to Actb and compared to M[0]. n = 3–4, representative of 3–5 experiments. Unpaired t test (*** p < 0.001). E) Schematic of timing variations for inhibitors and IL-4 polarization. Macrophages were treated with vehicle (DMSO), HC3 (250 μM), or RSM (5 μM) in the first 24 h then polarized with IL-4 (20 ng/mL) for 24 h (POST). Inhibitors were given together with IL-4 for 24 h (CONC). Macrophages were polarized with IL-4 for 24 h, then treated with inhibitors for 24 h (PRE). Expression of Retnla normalized to Actb and compared to M[0]. n = 3, representative of 1 experiment. Two-way ANOVA with Tukey’s test for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). F) Macrophages were treated with vehicle (DMSO), HC3 (250 μM) or RSM-932a (5 μM) for 24 h, washed, then treated with IL-4 (20 ng/mL) for 15’ and stained for intracellular pSTAT6 (Tyr641). n = 3, representative of 3 experiments. Two-way ANOVA with Tukey’s test for multiple comparisons (** p < 0.01, **** p < 0.0001). G) Surface PD-L1 and PD-L2 expression in M[0], M[IL-4], or M[LPS], treated with vehicle, HC3 (250 μM) or RSM-932a (5 μM). n = 2, representative of 3 experiments. H) Macrophages were treated with vehicle (DMSO) or HC3 (250 μM) for 24 h, washed, then treated with IL-4 (20 ng/mL) for 24 h in complete or choline-deficient (ΔCho) media. B) Quantification of intracellular RELMα staining in live F480 + macrophages. n = 3, representative of >4 experiments. One-way ANOVA with Tukey’s test for multiple comparisons (* p < 0.05, ** p < 0.01). I) Macrophages were treated with vehicle (DMSO), HC3 (250 μM), or RSM-932a (5 μM) for 24 h, washed, then treated with IL-4 (20 ng/mL) for 24 h. Detection of supernatant RELMα by ELISA. n = 5 (vehicle, HC3, IL-4) or n = 2 (RSM). Unpaired t test (** p < 0.01). Schematics were created using BioRender.

Article Snippet: Recombinant RELMα, polyclonal rabbit anti-RELMα capture, and polyclonal biotinylated rabbit anti-RELMα detection antibodies (all Peprotech) were used according to a previously described protocol [ ].

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: PLOS Pathogens

Article Title: Choline metabolism underpins macrophage IL-4 polarization and RELMα up-regulation in helminth infection

doi: 10.1371/journal.ppat.1011658

Figure Lengend Snippet: A) Schematic of 7-day in vivo choline kinase inhibition. Mice were treated intraperitoneally with vehicle (40% DMSO in PBS) or RSM-932a (3 mg/kg) every other day for 6 days and sacrificed on day 7. B) UMAP of peritoneal cell populations. n = 9–10, representing 2 independent experiments. C) Intracellular RELMα expression in live CD11b + F4/80 hi large (LPM) and CD11b + F4/80 lo small (SPM) peritoneal macrophages. Two-way ANOVA with Šídák’s test for multiple comparisons (* p < 0.05, **** p < 0.001). D-E) PD-L1 or PD-L2 expression (gMFI) in live CD11b + F4/80 hi LPM and CD11b + F4/80 lo SPM. Two-way ANOVA with Šídák’s test for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.001). F) Intracellular RELMα expression in LPM and SPM isolated from peritoneal cavity aggregates or suspended lavage cells. Two-way ANOVA with Šídák’s test for multiple comparisons (* p < 0.05, **** p < 0.0001). Schematics were created using BioRender.

Article Snippet: Recombinant RELMα, polyclonal rabbit anti-RELMα capture, and polyclonal biotinylated rabbit anti-RELMα detection antibodies (all Peprotech) were used according to a previously described protocol [ ].

Techniques: In Vivo, Inhibition, Expressing, Isolation

Journal: PLOS Pathogens

Article Title: Choline metabolism underpins macrophage IL-4 polarization and RELMα up-regulation in helminth infection

doi: 10.1371/journal.ppat.1011658

Figure Lengend Snippet: A) Schematic of primary infection with 200 H . polygyrus ( Hp ) L3 larvae through oral gavage. Mice were treated intraperitoneally with vehicle (40% DMSO in PBS) or RSM-932a (3 mg/kg) every other day for 8 days starting on day 8 and sacrificed on day 17. B-C) Percentage of weights at start of vehicle or RSM injections at 8 DPI with H . polygyrus (B) or final weights at day 17 (C). Mixed-effects analysis with Tukey’s test for multiple comparisons (* p < 0.05). D-E) Eggs in feces were counted at multiple time points after infection, and E) adult worms were isolated from the small intestine and enumerated on the day of sacrifice. Values represent means ± SEM (n = 3–5 mice per group), representative of 3 experiments. Two-way ANOVA with Šídák’s test for multiple comparisons and unpaired t test (ns). F-G) Detection of serum and G) peritoneal fluid RELMα by ELISA in naïve and H . polygyrus -infected mice. n = 3–5 per group. Two-way ANOVA with Šídák’s test for multiple comparisons (* p < 0.05, *** p < 0.001 for differences between treatment and ## p < 0.01 for differences between naïve and infected vehicle-treated mice). H-L) Immunofluorescent staining of intestinal tissue for CD206 and RELMα against DAPI counterstain. Scale bar 50 μm. Quantification of CD206 + (I) or RELMα + (J) per DAPI + cell. Unpaired t test (* p < 0.05). Quantification of F4/80 + (K) and RELMα + F4/80 + (L) per DAPI + cell. n = 6. Two-way ANOVA with Šídák’s test for multiple comparisons (**** p < 0.0001). Schematics were created using BioRender.

Article Snippet: Recombinant RELMα, polyclonal rabbit anti-RELMα capture, and polyclonal biotinylated rabbit anti-RELMα detection antibodies (all Peprotech) were used according to a previously described protocol [ ].

Techniques: Infection, Isolation, Enzyme-linked Immunosorbent Assay, Staining

Journal: Virology Journal

Article Title: Attenuation of vaccinia virus by the expression of human Flt3 ligand

doi: 10.1186/1743-422X-7-109

Figure Lengend Snippet: Incorporation of sFL into viral particles . Aliquots of the indicated sucrose purified virions (A) or their solubilized and separated fractions (B) were analyzed by western blot using the FL specific BAF308 monoclonal antibody. The sample prepared from the culture medium of 293T cells transfected with pBSC-FL expression plasmid served as a positive control. To neutralize virus infectivity, the saccharose purified particles were incubated at 37°C for 1 hour with PBS, anti-FL antibody or anti-VACV serum. The first aliquot of samples was used for infection of cell cultures (C) . Cytosine arabinoside (40 μg/ml) was added for inhibition of viral replication as the positive control. The beta-galactosidase assay was performed after 24 hours of cultivation. The second aliquot of samples was applied to a formvar membrane coated copper grid (D) . Negatively stained particles were examined under an electron-microscope at a magnification of 50 000×.

Article Snippet: FL was quantified with an Flt3 ligand ELISA detection kit (R&D Systems GmBH, Wiesbaden-Nordenstadt, Germany) using the capture mouse monoclonal antibody MAB608 (100 ng/well), biotinylated detection goat polyclonal antibody BAF308 (7.5 ng/well), streptavidin-HPR (1:250) or avidin-HPR (1:1000) complex, both obtained from Pharmingen (BD Biosciences, Erembodegem, Belgium), and TMB substrate solution for visualization of the reaction.

Techniques: Purification, Western Blot, Transfection, Expressing, Plasmid Preparation, Positive Control, Infection, Incubation, Inhibition, β-Gal Assay, Staining, Microscopy

Journal: BMC Pulmonary Medicine

Article Title: Plasma CC16 levels are associated with development of ALI/ARDS in patients with ventilator-associated pneumonia: a retrospective observational study

doi: 10.1186/1471-2466-9-49

Figure Lengend Snippet: Characteristics of the study population at study enrolment

Article Snippet: Then the plates were washed and incubated for one hour with 10 ng polyclonal biotinylated anti-human CC16 detection antibody A0257 (Dako, Glostrup, Denmark).

Techniques:

Journal: BMC Pulmonary Medicine

Article Title: Plasma CC16 levels are associated with development of ALI/ARDS in patients with ventilator-associated pneumonia: a retrospective observational study

doi: 10.1186/1471-2466-9-49

Figure Lengend Snippet: CC16 and sRAGE levels in VAP patients and control patients . Plasma levels of Clara cell protein (CC16) and soluble receptor for advanced glycation end products (sRAGE) in patients who developed ventilator-associated pneumonia (left graphs) and mechanically ventilated control patients (right graphs). In the left graphs day 0 represents the day of ventilator-associated pneumonia diagnosis. In the right graphs day 0 represents the day of start of mechanical ventilation.

Article Snippet: Then the plates were washed and incubated for one hour with 10 ng polyclonal biotinylated anti-human CC16 detection antibody A0257 (Dako, Glostrup, Denmark).

Techniques:

Journal: BMC Pulmonary Medicine

Article Title: Plasma CC16 levels are associated with development of ALI/ARDS in patients with ventilator-associated pneumonia: a retrospective observational study

doi: 10.1186/1471-2466-9-49

Figure Lengend Snippet: Biomarker levels in VAP patients with and without ALI/ARDS . Plasma levels of CC16, sRAGE, SP-D and KL-6 in patients developing ventilator-associated pneumonia. Open circles: patients without ALI/ARDS after onset of ventilator-associated pneumonia, closed circles: patients who progressed to ALI/ARDS at or after onset of ventilator-associated pneumonia.

Article Snippet: Then the plates were washed and incubated for one hour with 10 ng polyclonal biotinylated anti-human CC16 detection antibody A0257 (Dako, Glostrup, Denmark).

Techniques: Biomarker Assay

Journal: BMC Pulmonary Medicine

Article Title: Plasma CC16 levels are associated with development of ALI/ARDS in patients with ventilator-associated pneumonia: a retrospective observational study

doi: 10.1186/1471-2466-9-49

Figure Lengend Snippet: ROC curves . Receiver operating characteristic curves of CC16, sRAGE, SP-D and KL-6 for the diagnosis of ALI/ARDS on the day patients developed ventilator-associated pneumonia. AUC area under the curve.

Article Snippet: Then the plates were washed and incubated for one hour with 10 ng polyclonal biotinylated anti-human CC16 detection antibody A0257 (Dako, Glostrup, Denmark).

Techniques: