The segments of mouse brain were perfused and fixed with 4% paraformaldehyde in phosphate‐buffered saline (PBS) under physiological pressure and embedded in paraffin and sectioned at a thickness of 5 μm. Immunohistochemical analyses were performed using phospho‐SAPK/JNK (Thr183/Tyr185) antibody (#9251S; Cell Signaling Technology), anti‐GFAP antibody (Z0334; Dako), anti‐S100 beta antibody [EP1576Y] (ab52642; Abcam), and anti‐beta amyloid 1–42 antibody (ab14220; Abcam). The deparaffinized and dehydrated sections were incubated for 5 min at 120 °C in Dako Target Retrieval Solution (pH 9.0; S2367; Dako), treated with in 3% hydrogen peroxide in distilled water (dH2O), and incubated with phospho‐SAPK/JNK (Thr183/Tyr185) antibody diluted 1:100 in TBS [0.02 m TRIZMA BASE, 0.137 m NaCl (pH 7.6)], anti‐GFAP antibody (Z0334; Dako) diluted 1:500 in TBS, anti‐S100 beta antibody [EP1576Y] diluted 1:500 in TBS, and anti‐beta amyloid 1–42 antibody (ab14220; Abcam) diluted 1:500 in TBS in each reaction after blocking nonspecific protein binding in TBS containing 5% goat serum for 10 min. Sections were treated with Histofine® Simple Stain™ Mouse MAX PO (R) (414341F; Nichirei Biosciences Inc., Tokyo, Japan) after washing in TBS and treated in diaminobenzidine and hematoxylin nuclear staining.
Ab14220
Manufactured by Abcam
Sourced in United States
Ab14220 is a mouse monoclonal antibody that targets the TGF-beta 1 protein. It is suitable for use in various laboratory applications such as Western blotting, ELISA, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Ab52642, by Abcam (1 mentions)
S2367, by Agilent Technologies (1 mentions)
Z0334, by Agilent Technologies (1 mentions)
Histofine simple stain mouse max po r, by Nichirei Biosciences (1 mentions)
Ab1903, by Abcam (1 mentions)
Avidin biotin peroxidase complex, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine tetrahydrochloride dab, by Merck Group (1 mentions)
2 protocols using ab14220
1
Immunohistochemical Analysis of Mouse Brain
2
Immunohistochemical Analysis of Protein Aggregation
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Imunohistochemical analysis of protein aggregation was evaluated in three slices/region/animal, with a total of 15 slices/region/animal taken, between Bregma levels from -3.6 mm to -4.1 mm for the hippocampus, -5.5 mm to -5.8 mm for the substantia nigra and -9.6 to -10.0 for the locus coeruleus. The whole hippocampus, substantia nigra and locus coeruleus were analyzed, without any distinction of the subarea (hippocampus: CA/dentate gyrus; substantia nigra: pars compacta/pars reticulata)
Sections were incubated for 24 hours at 4°C with antibodies against either hyperphosphorylated TAU 1/200 (Santa Cruz Biotechnology, sc-101813), alpha-synuclein 1/200 (Abcam, ab1903) or amyloid-beta peptide 1/200 (Abcam, ab14220), followed by incubation with biotinylated secondary goat anti-rabbit immunoglobulin antibodies (1/230, Vector, USA) for 90 min. Sections were then incubated with avidin-biotin peroxidase complex (1:120, Vectastain, Vector, USA) for 45 minutes and immunoreactivity was visualized using 3-3'-diaminobenzidine tetrahydrochloride (DAB, Sigma) as chromogen and H 2 O 2 (0.05%) for six minutes of reaction. Cells were counterstained with hematoxylin, which is a valuable tool to stain nuclei. The use of hematoxylin facilitates intracellular localization of protein aggregates.
Sections were incubated for 24 hours at 4°C with antibodies against either hyperphosphorylated TAU 1/200 (Santa Cruz Biotechnology, sc-101813), alpha-synuclein 1/200 (Abcam, ab1903) or amyloid-beta peptide 1/200 (Abcam, ab14220), followed by incubation with biotinylated secondary goat anti-rabbit immunoglobulin antibodies (1/230, Vector, USA) for 90 min. Sections were then incubated with avidin-biotin peroxidase complex (1:120, Vectastain, Vector, USA) for 45 minutes and immunoreactivity was visualized using 3-3'-diaminobenzidine tetrahydrochloride (DAB, Sigma) as chromogen and H 2 O 2 (0.05%) for six minutes of reaction. Cells were counterstained with hematoxylin, which is a valuable tool to stain nuclei. The use of hematoxylin facilitates intracellular localization of protein aggregates.
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