THP1 cells were seeded on cover slips and differentiated into macrophage (THP1-MΦ), fixed in 4% paraformaldehyde (PFA, 10 min at rt), washed (1 X PBST, thrice 5 min each), permeabilized with 0.1% triton X100 (15 min, 1 X PBST, rt), and blocked with 3% BSA (in 1XPBST for 1 h). The cells were then incubated with mouse anti-human CD68 primary antibody (# 14-0688-82, Invitrogen, 4 h, rt). Cells were washed 3 times with PBST followed by incubation with FITC-conjugated goat Anti-Mouse (# 6785, Abcam) secondary antibodies for 1 h at RT. DAPI was added to the cells with a final concentration of 1 μg/mL and incubated for 15 min. Finally, the cells were washed 3 times with PBST and fixed with mounting media on a slide and analyzed under a fluorescence microscope (Nikon ECLIPSE TE2000-U).
Fitc conjugated goat anti mouse
Manufactured by Abcam
FITC-conjugated goat Anti-Mouse is a secondary antibody conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate). It is designed to specifically bind to mouse primary antibodies, allowing for the detection and visualization of target proteins or antigens in various immunoassays and imaging techniques.
Automatically generated - may contain errors
2 protocols using fitc conjugated goat anti mouse
1
Immunofluorescence Staining of THP-1 Macrophages
2
Immunohistochemical Analysis of IPF Lung Tissue
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Lung tissue sections obtained from IPF were deparaffinized and rehydrated in increasing concentrations of ethanol. The sections were incubated with 5% bovine serum albumin (BSA) in TBS for 1 h to block non-specific binding of antibodies and then with a mixture of polyclonal anti-S100A4 (1:300, Abcam) and monoclonal anti-αSMA (1:300, Abcam) primary antibodies in 5% BSA in TBS overnight at 4 � C. After washing with 1 � TBS, the sections were incubated with a mixture of fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse (1:2000, Abcam) and phycoerythrin (PE)-conjugated donkey anti-rabbit (1:2000, Abcam) secondary antibodies. Nuclei were counterstained with 4 0 ,6diamidino-2-phenylindole (Invitrogen, Carlsbad, CA). Confocal laser scanning microscopy was performed using a 510 META LSM ( � 40 objective) coupled to a Coolsnap Photometrics HQ camera (Photometrics, AZ), and images were generated using the Zeiss LSM image browser.
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