To determine if the NADPH oxidase complex, specific, and azurophilic granules are recruited to the L. pneumophila phagosome, PMNs were plated into 24-well plates containing glass coverslips (3 × 106 cells per well) and infected with either post-exponential-phase wild type, the ΔT4SS and ΔlamA mutants, or the complemented lamA/C mutant at an MOI of 1. Additionally, killed and opsonized S. aureus was used as a positive control for marker recruitment. At 5, 15, and 60 min postinfection, the monolayers were fixed with 4% formaldehyde for 30 min and permeabilized with 0.1% Triton X-100 in PBS for 15 min. The monolayers were labeled with rabbit anti-Legionella antiserum (1:1,000 dilution) in conjunction with mouse anti-p47phox (BD Transduction Laboratories), anti-lactoferrin (Abcam), or anti-elastase (Abcam) antibodies. PMNs were infected with bacteria and analyzed by confocal microscopy.
Anti lactoferrin
Manufactured by Abcam
Sourced in United States
Anti-lactoferrin is a laboratory reagent that can be used to detect and quantify the presence of the lactoferrin protein in biological samples. Lactoferrin is an iron-binding glycoprotein found in various bodily fluids and is involved in immune response and iron homeostasis. Anti-lactoferrin can be utilized in various research and diagnostic applications.
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Lab products found in correlation
2 protocols using anti lactoferrin
1
Recruitment of Phagocyte Granules to Legionella
2
Quantifying Lactoferrin Levels via ELISA
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The assay was carried out as formerly described [33 (link)]. Costar ELISA plate (Cambridge, USA) was coated with 50 μL of each in vitro reaction. After an incubation period of 24 h at 4 °C; the plate was washed five times with 0.12 M NaCl, 0.04 M sodium phosphate pH 7.2 buffer (PBS), and then 100 μL of blocking buffer (2% gelatin in PBS) were added for 1 h at 37 °C. Then 50 μL of rabbit polyclonal anti-lactoferrin (Abcam, Cambridge, MA, USA) diluted 1:100 in 2% gelatin-PBS were added to each well. After 1 h of incubation at 37 °C, the plate was washed five times with PBS, and 50 μL of alkaline phosphatase–conjugated anti-rabbit IgG (BIO-RAD, Alfred Nobel, Hercules, USA) diluted 1:1000 with 2% gelatin–PBS were added, followed by an incubation of 1 h at 37 °C. p-Nitrophenyl phosphate (p-NPP) was added for color development, and optical density was measured at 405 nm.
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