Brains were quickly removed after decapitation. Striatum and midbrain samples were obtained at 0 °C and subsequently homogenized using Disposable Pellet Mixers with Pestle Motor (VWR). Tissue homogenates were prepared in RIPA buffer; pH 7.5, containing 10% Triton 500 μl, 5 μl aprotinin, PMSF 50 μl, Na3VO4 100 μl and NaF 20 μl in 4.32 ml PBS. Extracts were clarified by centrifugation at 4 °C (13,200g for 20 min). Supernatants were collected and eluted with RIPA buffer, and the proteins were resolved by SDS-PAGE [45 (link)]. The primary antibodies used were: β-Tubulin (#05-661, Millipore), β-Actin (Abcam, ab6276, AC15), HA-Tag (C29F4) (#3724, Cell Signalling), Tyrosine Hydroxylase (MAB318, Sigma Aldrich), α-synuclein (610787, BD Transduction Laboratories™), DAT (MAB369, Merckmillipore), LC3 (5F10, Nanotools Antibodies), Tom20 (D8T4N) (#42406, Cell Signalling), Mitofusin-2 (D2D10) (#9482, Cell Signalling), DRP1 (D6C7) (#8570, Cell Signalling), Phospho-Tau Ser202/Thr205 (AT8) (#MN1020, ThermoFisher), and Dopamine D2 Receptor (AB5084P, Merckmillipore). The rabbit anti-mouse (GE healthcare UK, NA934V), or mouse anti-rabbit (GE health UK, NA931V) were used to react with the corresponding primary antibodies. Immunoreactive bands were visualized by enhanced chemiluminescence (GE Healthcare). Densitometry analysis on the bands was calculated using ImageJ software.
Disposable pellet mixers with pestle motor
Manufactured by Avantor
Disposable Pellet Mixers with Pestle Motor are a type of laboratory equipment used for mixing and homogenizing samples. The device consists of a disposable mixing chamber and a motorized pestle that can be used to thoroughly mix various materials, such as powders, liquids, or suspensions.
Automatically generated - may contain errors
Lab products found in correlation
Tyrosine hydroxylase, by Merck Group (2 mentions)
Enhanced chemiluminescence, by GE Healthcare (2 mentions)
β tubulin, by Merck Group (2 mentions)
Drp1 d6c7, by Cell Signaling Technology (2 mentions)
Tom20 d8t4n, by Cell Signaling Technology (2 mentions)
Dopamine d2 receptor, by Merck Group (2 mentions)
Phospho tau ser202 thr205 at8, by Thermo Fisher Scientific (2 mentions)
Mitofusin 2 d2d10, by Cell Signaling Technology (2 mentions)
Ha tag c29f4, by Cell Signaling Technology (2 mentions)
β actin, by Abcam (2 mentions)
α synuclein, by BD (1 mentions)
2 protocols using disposable pellet mixers with pestle motor
1
Protein Extraction and Western Blot Analysis of Neural Tissues
2
Protein Extraction and Analysis from Brain Tissues
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Brains were quickly removed after decapitation. Striatum and midbrain samples were obtained at 0°C and subsequently homogenized using Disposable Pellet Mixers with Pestle Motor (VWR). Tissue homogenates were prepared in RIPA buffer; pH 7.5, containing 10% Triton 500μl, 5μl aprotinin, PMSF 50μl, Na 3 VO 4 100μl and NaF 20μl in 4.32 ml PBS. Extracts were clari ed by centrifugation at 4°C (13,200 g for 20 minutes). Supernatants were collected and eluted with RIPA buffer, and the proteins were resolved by SDS-PAGE (43) . The primary antibodies used were: β-Tubulin (#05-661, Millipore), β-Actin (Abcam, ab6276, AC15), HA-Tag (C29F4) (#3724, Cell Signalling), Tyrosine Hydroxylase (MAB318, Sigma Aldrich), a-synuclein (610787, BD Transduction Laboratories™), DAT (MAB369, Merckmillipore), LC3 (5F10, Nanotools Antibodies), Tom20 (D8T4N) (#42406, Cell Signalling), Mitofusin-2 (D2D10) (#9482, Cell Signalling), DRP1 (D6C7) (#8570, Cell Signalling), Phospho-Tau Ser202/Thr205 (AT8) (#MN1020, ThermoFisher), and Dopamine D2 Receptor (AB5084P, Merckmillipore). The rabbit anti-mouse (GE healthcare UK, NA934V), or mouse anti-rabbit (GE health UK, NA931V) were used to react with the corresponding primary antibodies.
Immunoreactive bands were visualized by enhanced chemiluminescence (GE Healthcare). Densitometry analysis on the bands was calculated using ImageJ software.
Immunoreactive bands were visualized by enhanced chemiluminescence (GE Healthcare). Densitometry analysis on the bands was calculated using ImageJ software.
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