Microtubule-associated protein 2 (MAP-2) immunostaining was performed to evaluate brain tissue loss. At 21 days after surgery, mice (n = 6 per group) were anesthetized with pentobarbital sodium (50 mg/kg, i.p.) and transcardially perfused with 0.9% saline. Brains were removed and fixed with 4% paraformaldehyde (PFA) overnight at 4°C and then dehydrated in 20% to 30% sucrose solutions. Serial coronal sections (20 μm) were cut with a freezing microtome (Leica, Germany). The sections were washed in phosphate-buffered saline (PBS) and incubated with 10% normal goat serum (ZSGB-BIO, China) for 60 minutes at room temperature. The sections were then incubated with rabbit anti-MAP-2 antibody (1 : 1000, AB221693, Abcam) overnight at 4°C. After washing with PBST, the samples were further incubated with donkey anti-rabbit IgG-CFL 488 (1 : 100, sc-362261, Santa Cruz) for 2 hours at room temperature. All samples were examined under a laser confocal microscope (Nikon, Japan). The analysis and calculation method of brain tissue loss were consistent with the measurement method of infarct volume.
Ab221693
Manufactured by Abcam
AB221693 is a polyclonal antibody that recognizes the human Epidermal Growth Factor Receptor (EGFR) protein. This antibody is intended for use in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Normal goat serum (ngs), by ZSGB-BIO (1 mentions)
Laser confocal microscope, by Nikon (1 mentions)
Donkey anti rabbit igg cfl 488, by Santa Cruz Biotechnology (1 mentions)
Freezing microtome, by Leica (1 mentions)
Ab33922, by Abcam (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Ab6785, by Abcam (1 mentions)
2 protocols using ab221693
1
Evaluating Brain Tissue Loss via MAP-2 Immunostaining
2
Immunofluorescence Staining of Neurons and Astrocytes
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The neurons and astrocytes were washed with PBS three times and fixed with 4% paraformaldehyde for 10 min. Then, the cells were washed with PBS three times and permeabilized with Triton for 10 min. Following PBS washing three times, the cells were blocked with 0.5% bovine serum albumin for 30 min and cultured with the primary antibodies GFAP (ab33922, 1:2000, Abcam) and MAP2 (ab221693, 1:1000, Abcam) at 4℃ overnight. On the next day, the cells were cultured at room temperature for 15 min, washed with PBS three times, and cultured with the secondary antibody fluorescein isothiocyanate (ab6785, 1:10000, Abcam) in the dark at 37℃ for 90 min. Following PBS washing three times, the cells were sealed with fluorescent mounting media and observed under the fluorescence microscope (Olympus, Tokyo, Japan).
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