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Ab1793

Manufactured by AbFrontier

AB1793 is a compact and versatile laboratory centrifuge designed for a wide range of applications. It features a high-speed motor and a robust construction to ensure reliable and efficient performance. The product specifications and details are available upon request.

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4 protocols using ab1793

1

Western Blot Analysis of Lung Tissue

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For western blot analysis, proteins collected from whole lung tissue were separated by SDS-PAGE in a 15% polyacrylamide gel for 3 h. The separated proteins were then transferred to PVDF membranes (Bio-Rad, USA) for 1 h at 90 volts. After blocking the membranes overnight at 4 °C with 5% skim milk, they were probed using anti-GAPDH (Santa Cruz Biotechnology, LF-PA0018), polyclonal mouse anti-NF-κB (Santa Cruz Biotechnology, SC-71675), monoclonal mouse anti-TNF-α (ABfrontier, AB1793), polyclonal mouse anti-IL-1β (ABfrontier, AB1413), polyclonal mouse anti-Cathelicidin (ABfrontier, AB93357) and polyclonal mouse anti-PGC-1 (Millipore AB3242). The membranes were then washed with TBST buffer and incubated with secondary antibodies (goat anti-mouse IgG (HRP) LF-SA5001-conjugated). Finally, the blots were developed using a western blot detection kit (LF-QC0103, Abfrontier)74 (link).
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2

Immunohistochemical Analysis of Lung and Visceral Fat

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Lungs and visceral fat were extracted from mice, washed once with phosphate buffered saline (PBS) and fixed in 4% paraformaldehyde for 24 h at 4 °C. The fixed tissue was then dehydrated through a 50–100% ethanol series (2 h at each step) and 3 incubations (1 h each) in xylene, embedded in paraffin and cut into 4-μm-thick sections using a microtome. Sections were incubated for 30 min at room temperature with primary antibody in 5% bovine serum albumin (BSA). The primary antibodies used were polyclonal mouse anti-cathelicidin (ABfrontier, AB93357), monoclonal mouse anti-TNF-α (ABfrontier, AB1793) and polyclonal mouse anti-IL-1β (ABfrontier, AB1413). The samples were washed with tris-buffered saline containing tween (TBST buffer), incubated with secondary antibody (goat anti-mouse IgG (HRP) LF-SA5001-conjugated), and stained with hematoxylin and eosin (H&E). Stained sections were examined under a fluorescence microscope (Ix71, Olympus, Tokyo, Japan)64 (link).
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3

Analyzing RAW 264.7 Cells' Cytokine Response to Staphylococcus aureus LTA

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RAW 264.7 cells (1 × 106 cells/mL) were cultured for 12 h as described above and incubated for 6 h in the presence of S. aureus LTA (1 μg/mL) with or without CSP-4 (100 μg/mL). The cells were then washed in PBS, fixed in 4% paraformaldehyde (15 min), permeabilized using 0.5% Triton-X 100 diluted in PBS, washed again 3 times by gently shaking in PBS for 5 min each, blocked in a blocking buffer (5% bovine serum albumin in PBS), and then added to an 8-well plate (200 μL/well), in which they were incubated for 1 h with gentle shaking. Primary antibodies (TNF-α, AbFrontier, Seoul, Korea, AB1793; and IL-1β AbFrontier, AB1413; 1:50 dilution) were then added to the cells for 24 h at 4° C, after which the cells were washed 3 times in PBS (5 min/wash). A secondary antibody was applied (goat anti-mouse IgG-HRP, LF-SA 5001-conjugated; 1:200 dilution) for 1 h at 37° C, and nuclei were stained using DAPI (0.25 µg/mL). The staining was visualized under a confocal laser-scanning microscope (LSM510; Carl Zeiss; Gottingen, Germany) [45 (link)].
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4

Evaluating CSP-4 Peptide in S. aureus Infection

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Male hairless mouse (aged 6 weeks, 5 mouse/group, total 7 groups) were injected intraepidermal with S. aureus CCARM 0027 (1 × 108 cfu/ml). The CSP-4 peptide (200, 100, 50, 25 μg/ml) was injected intraeptidermal into mice at 1 h after the S. aureus CCARM 0027 injection, samples of inflamed skin was collected after 7 days and 4-μm-thick sections were prepared as described in the preceding section. Each sample was then incubated for 30 min at 24° C with 5% bovine serum albumin, mouse anti-Toll-like receptor-2 (TLR-2) (AbFrontier, AB24192), mouse monoclonal anti-TNF-α (AbFrontier, AB1793), and mouse polyclonal anti-IL-1β (AbFrontier, AB1413). The samples were then washed with Tris-buffered saline containing Tween 20 buffer and incubated with HRP LF-SA5001-conjugated goat anti-mouse IgG, and stained with hematoxylin and eosin. The stained sections were examined under a fluorescence microscope [46 (link)].
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