To validate in vivo imaging data mice from each experimental group were euthanized by overdose of anesthetic. For the double-immunofluorescence analysis, mice were perfused with PBS (pH 7.5) followed by 40 mg/ml paraformaldehyde and incubated overnight in 200 mg/ml phosphate-buffered sucrose. The sections were blocked in 10% goat serum and then incubated overnight at room temperature using primary antibodies diluted in 5% goat serum/PBS1x+0.25% Triton X-100 (1:500 rabbit polyclonal anti-Iba1 (Wako), 1:500 anti-Gal-3 (ATCC), 1:250 rabbit anti IL-4 receptor (Santa Cruz), 1:500 rabbit anti CD11b (Serotech), 1:500 rabbit polyclonal anti-Ym1 (Stem Cell technologies) and 1:500 rabbit anti actin (Invitrogen). After washes in PBS, the sections were then incubated in corresponding fluorescent goat secondary antiserum for 2 h (1:500, Invitrogen), washed and mounted with Flouromount G (Thermofisher) (Rahimian et al., 2019b (link)). Alexa Fluor® 488 phalloidin (Thermo Fisher Scientific) was used to study microglia morphology (Leica CTR 500 microscope).
Rabbit anti actin
Manufactured by Thermo Fisher Scientific
Rabbit anti actin is a primary antibody that recognizes the actin protein, a fundamental component of the cytoskeleton in eukaryotic cells. It is commonly used in laboratory applications for the detection and analysis of actin in various cell and tissue samples.
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Lab products found in correlation
Flouromount g, by Thermo Fisher Scientific (1 mentions)
Anti ym1, by STEMCELL (1 mentions)
Rabbit anti il 4 receptor, by Santa Cruz Biotechnology (1 mentions)
Anti iba1, by Fujifilm (1 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions)
Anti rabbit igg microbeads, by Miltenyi Biotec (1 mentions)
Ls column, by Miltenyi Biotec (1 mentions)
2 protocols using rabbit anti actin
1
Immunofluorescent Staining of Microglia
2
Yeast Display Library Induction and Sorting
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To induce the SNU475 yeast display library, the cell population grown in SD-CAA medium was moved to SG-CAA medium (SD-CAA with 2% galactose instead of dextrose) and incubated at 20°C for 40 h. In 100 µl of MACS buffer (PBSM; 0.8% NaCl, 0.02% KCl, 0.144% Na 2 HPO 4 , 0.024% KH 2 PO 4 , 0.5% BSA, and 0.0744% EDTA-2Na), 3 × 10 7 induced cells were added and labeled using 5 µg of polyclonal rabbit anti-actin (Invitrogen) at RT for 30 min. After washing with PBSM, 20 µl of anti-rabbit IgG Microbeads (MiltenyiBiotec GmbH, Gladbach, Germany) and 80 µl of resuspended cells were incubated at 4°C for 30 min. After washing again, 3 × 10 7 cells in 500 µl of PBSM were loaded onto the LS column (MiltenyiBiotec GmbH, Gladbach, Germany) surrounded by magnet and washed with 10 ml of PBSM. MAC-sorted cells were incubated in 4 ml of SD-CAA medium with 100 µl of YPD at 30°C for 2 days [4] .
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