polyclonal goat igg af154  (R&D Systems)

Journal: JID Innovations

Article Title: Human keratinocytes exhibit limited potential for SARS-CoV-2 infection despite ACE2 and mature cathepsin L expression

doi: 10.1016/j.xjidi.2025.100447

Figure Lengend Snippet: Differential ACE2, TMPRSS2, NRP1, CTSL, CD147, AXL, and DPP4 protein expression. Whole-cell lysates of the indicated cells were analyzed by western blot using the following antibodies: ACE2 (mAb clone AC384), TMPRSS2 (mAb clone S20014A), NRP1 (mAb clone 14H4), CTSL (mAb clone 33/1), CD147 (mAb clone HIM6), AXL (polyclonal goat IgG, AF154), and DPP4 (polyclonal goat IgG, AF1180). ( a ) Representative immunoblots from 2 to 4 independent experiments. ( b ) Densitometric quantification normalized to actin, expressed as mean ± SEM from 2 to 4 independent experiments. CTSL, cathepsin L.

Article Snippet: Primary antibodies included anti-ACE2, clone AC384 (AdipoGen Life Science/Coger, Paris, France); anti-TMPRSS2, clone S20014A; anti-NRP1, clone 14H4; anti-CD147, clone HIM6; anti–keratin 10, rabbit polyclonal Poly19054 (BioLegend); anti-CTSL, clone 33/1 (eBioscience, Thermo Fisher Scientific); anti-AXL, polyclonal goat IgG AF154; and anti-DPP4, polyclonal goat IgG AF1180 (R&D Systems, Bio-Techne SAS, Noyal Châtillon, France).

Techniques: Expressing, Western Blot

Journal: Virology

Article Title: T-cell immunoglobulin and mucin (TIM) contributes to the infection of human airway epithelial cells by pseudotype viruses containing Hantaan virus glycoproteins.

doi: 10.1016/j.virol.2020.02.002

Figure Lengend Snippet: Fig. 5. TIM-1, αvβ3 integrin, and heparan sulfate can contribute to cell infection of HTNV pseudoviruses. (A) Expression of αvβ3 integrin in A549 cells. Live non-permeabilised cells were stained with the mouse anti-αvβ3 integrin mAb (red line). (B) Anti-αvβ3 integrin Ab reduces infection of VSV*ΔG-Luc(HTNV-G) but not VSV*ΔG-Luc(ANDV-G) into A549 cells. A549 cells were pre-treated for 1 h at 4 °C with mouse anti-αvβ3 mAb and a control mouse at 30 nM. Cells were then infected with VSV*ΔG-Luc(HTNV-G), VSV*ΔG-Luc(ANDV-G) and VSV*ΔG-Luc(EBOV-G), and shifted to 37 °C. Infection was quantified by counting EGFP-positive infected cells per well. (C) Relative contribution of αvβ3 integrin and hTIM-1. A549 cells were pre-treated with anti-αvβ3 and anti-hTIM-1 antibodies at 30 nM. The ex- periment settings were identical to the antibody perturbation assay as in panel (B). (D) Heparan sulfate reduces infection of HTNV but not ANDV pseudotypes into A549 cells. VSV*ΔG-Luc(HTNV-G), VSV*ΔG-Luc(ANDV-G), VSV*ΔG-Luc(EBOV-G), VSV*ΔG-Luc(LASV-G) and AdV5-GFP were pre-incubated with soluble heparan sulfate at various concentrations and used to infect monolayers of A549 cells. Infection levels were assessed by counting EGFP-positive infected cells. (E) Heparan sulfate and anti-hTIM-1 block HTNV-G infection. A549 cells were pre-treated for 1 h at 4 °C with the polyclonal Ab goat anti-hTIM-1 at 30 nM. Meantime, VSV*ΔG- Luc(HTNV-G), was pre-incubated with heparan sulfate (HS) at 5 μg/ml. Cells were then infected with pre-treated VSV*ΔG-Luc(HTNV-G) and shifted to 37 °C. Infection was quantified 16 h post-infection by counting EGFP-positive infected cells. (F) Relative contribution of αvβ3 integrin and heparan sulfate. Pre-treatment of cells with anti-αvβ3 mAb (30 nM), viruses with 5 μg/ml of heparan sulfate (HS) and infections were assessed as in Fig. 5E. Infection was quantified 16 h post-infection by counting EGFP-positive infected cells. (B–F) Data are means + SD (n = 3) with p-value *: p ≤0.05; **: p ≤0.01; ***: p ≤0.001.

Article Snippet: Human TIMD4 was obtained from Sino Biological Inc. Purified goat anti-human Axl IgG polyclonal antibody (pAb, #AF154), purified goat anti-human TIM-1/KIM-1/HACVR IgG pAb (#AF1750), phycoerythrin (PE)-conjugated donkey anti-goat IgG pAb, purified donkey anti-goat IgG pAb (#AF109) and monoclonal mouse IgG (#MAB002) were from R&D Systems.

Techniques: Infection, Expressing, Staining, Control, Incubation, Blocking Assay