BV2 microglia were grown on coverslips in 24-well plates. After fixation in 4% paraformaldehyde for 15 min, the cells were blocked with goat serum (ZsBio, China) for 20 min at room temperature and then incubated at 37 °C for 1 h with rabbit anti-mouse Arg1 (Millipore, USA), rabbit anti-human CD206 (Santa Cruz, USA) and mouse anti-mouse phospho-AKT (CST, USA). After the slides were washed three times, they were incubated at 37 °C for 1 h in the dark with a secondary fluorescent chicken anti-rabbit IgG (H + L) CF633 (Sigma-Aldrich, USA) and rabbit anti-mouse AlexaFluor 488 (Lifetech, USA). After the slides were washed, the cells were stained with DAPI (Beyotime Biotechnology, China) for 15 min at 37 °C, and then the slides were mounted with an anti-fluorescence quenching agent. Images were visualized using an LSM 780 confocal laser scanning microscope (Carl Zeiss GmbH, Germany).
Rabbit anti mouse alexafluor 488
Manufactured by Thermo Fisher Scientific
Sourced in United States
Rabbit anti-mouse AlexaFluor 488 is a secondary antibody that binds to mouse primary antibodies and is conjugated to the AlexaFluor 488 fluorescent dye. It is used in immunofluorescence and other fluorescence-based applications to detect and visualize target proteins or molecules.
Automatically generated - may contain errors
Lab products found in correlation
Goat serum, by ZSGB-BIO (2 mentions)
Lsm 780 laser scanning confocal microscope, by Zeiss (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Alexa fluor 568 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Axio imager z1 fluorescent microscope, by Zeiss (1 mentions)
Bx60 fluorescence microscope, by Olympus (1 mentions)
5 protocols using rabbit anti mouse alexafluor 488
1
Immunofluorescent Staining of BV2 Microglia
2
Immunofluorescence Imaging of TREM2, Akt, and CD206
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Cells were grown on coverslips in 24-well plates. After being fixed and permeabilized, the cells were blocked with goat serum (ZsBio) for 20 min at room temperature and washed three times in PBS. The cells were incubated with rat anti-mouse TREM2 (Merck Millipore), rabbit anti-mouse phosphor-Ser473-Akt (Cell Signaling Technology, Danvers, MA, USA), and/or mouse anti-mouse CD206 (Santa Cruz) antibodies at 37°C for 1 h. After being washed, the slides were incubated for 1 h at 37 °C with rabbit anti-mouse AlexaFluor 488 (Life Technologies, Carlsbad, CA, USA), goat anti-rat CF568 (Sigma–Aldrich, St. Louis, MO, USA), and chicken anti-rabbit CF633 antibodies (Sigma–Aldrich, St. Louis, MO, USA) in the dark. The slides were then washed and mounted with an aqueous-based anti-fade mounting medium. Images of stained cells were captured using an LSM 780 confocal laser-scanning microscope (Carl Zeiss GmbH, Jena, Germany).
3
Immunofluorescence Analysis of PAX8 and Calretinin
4
Measuring TRAIL Receptor Expression
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Cells were incubated with different concentrations of ATO for 4 to 48 hs, harvested by trypsinization and fixed by ethanol. Cells were incubated with antibodies in staining buffer (1% bovine serum albumin and 0.2% (v/v) Triton-X in PBS) for 20 min and were subsequently analyzed using the FACSCanto II cytometer. The following antibodies were used for staining: mouse anti-TRAIL-R1 (clone DJR1) and TRAIL-R2 (clone DJR2-4, both PE-labeled, eBioscience, Hatfield, UK); mouse anti-p-H2AX (clone JBW301, Merck-Millipore, Darmstadt, Germany), and rabbit anti-mouse Alexa Fluor 488 (Life Technologies, Darmstadt, Germany).
5
Immunofluorescence Microscopy of α3β1 and LAMP1
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Cells were grown in 8-well chamber Lab-Tek Pernox slides. They were fixed with 4% paraformaldehyde-lysine-periodate (49 (link)) treated with 1% SDS in PBS for antigen retrieval and blocked in 1% bovine serum albumin in PBS supplemented with 0.2% Triton X-100. The α3 subunit was stained with a goat polyclonal antibody (Santa Cruz, C16) followed by donkey-anti-goat Alexa-fluor 555. The β1 subunit was stained by mouse monoclonal antibody M17-P5-F11 followed by rabbit-anti mouse Alexa-fluor 488 (Life Technologies). LAMP1 was stained with rabbit mAb D2D1 (9091) (Cell Signaling Technology), followed by goat-anti-rabbit Alexa-Fluor 555. Images were obtained on an Olympus BX60 fluorescence microscope.
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