Ab152

Manufactured by Merck Group

Sourced in United States, Germany, Canada, United Kingdom, Japan, China

The AB152 is a laboratory centrifuge designed for general-purpose applications. It can separate various biological samples, such as cells, proteins, and DNA, based on their density differences. The centrifuge operates at a maximum speed of 15,000 RPM and can accommodate a variety of rotor configurations to meet the needs of different research and diagnostic applications.

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Lab products found in correlation

Ab13970, by Abcam (11 mentions) Alexa fluor 488, by Thermo Fisher Scientific (10 mentions) Mab369, by Merck Group (9 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (9 mentions) Mab318, by Merck Group (7 mentions) Lsm 710, by Zeiss (6 mentions) 49 6 diamidino 2 phenylindole dapi, by Vector Laboratories (6 mentions) Mab377, by Merck Group (5 mentions) Mab5406, by Merck Group (5 mentions) Cyanine3 (cy3), by Jackson ImmunoResearch (4 mentions) Ab51253, by Abcam (4 mentions) Alexa fluor 594, by Thermo Fisher Scientific (4 mentions) Pk 6100, by Vector Laboratories (4 mentions) Ab144p, by Merck Group (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (4 mentions) Mab360, by Merck Group (4 mentions) Ab1542, by Merck Group (4 mentions) A1r confocal microscope, by Nikon (3 mentions) Fv1000, by Olympus (3 mentions)

243 protocols using ab152

Immunohistochemical analysis of Parkinson's disease

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Procedures for the preparation of brain tissues and immunohistochemistry are described in the Supplementary Methods (Additional file 1).
Primary antibodies (immunofluorescence): anti-TH (Millipore #AB152) 1:1000; anti-α-syn (Millipore #AB5334P) 1:1000; anti-α-syn clone 5G4 (AJ Roboscreen) 1:1000; anti-phospho S129 α-syn (Abcam ab59264) 1:1000. Secondary antibodies: Cy2-conjugated donkey anti-rabbit IgG (H + L) (Jackson ImmunoResearch Inc. #711–225-152) 1:1000; Cy3-conjugated F(ab’)₂ fragment donkey anti-sheep IgG (H + L) (Jackson ImmunoResearch Inc. #713–166-147) 1:1000.
Primary antibodies (DAB): anti-TH (Millipore #AB152 1:1000); anti-DAT (Millipore #MAB369) 1:4000; anti-HA-tag (Covance clone 16B12 #MMS-101P) 1:1000. Secondary antibodies: peroxidase Goat anti-rabbit IgG (Vector Laboratories #PI-1000) 1:200; peroxidase goat anti-mouse IgG (Vector Laboratories #BA-9200) 1:200; biotinylated rabbit anti-rat IgG (Vector Laboratories #BA-4001).

Bobela W., Nazeeruddin S., Knott G., Aebischer P, & Schneider B.L. (2017). Modulating the catalytic activity of AMPK has neuroprotective effects against α-synuclein toxicity. Molecular Neurodegeneration, 12, 80.

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Quantitative Immunohistochemical Analysis of Dopaminergic Neurons

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Mice were anesthetized and perfused with 4% paraformaldehyde. Brains were extracted, dehydrated in 30% sucrose solution, embedded in OCT, and continuously sliced at 25 μm thickness. The immunohistochemistry staining was processed as previously described³¹ (link): tissue sections were blocked with 3% H₂O₂ for 15 min and 5% goat serum for 30 min at room temperature after heat-induced antigen retrieval. The sections were then incubated with primary antibody to TH (1:500, AB152, Merck-Millipore) at 4 °C for 12 h and followed by incubating with horseradish peroxidase (HRP)-labeled goat anti-rabbit (1:300; 31460, ThermoFisher Scientific). The number of TH-positive cells in the SN area was determined by stereological methods, as previously described³² (link). For immunofluorescence staining: tissue sections were blocked with 5% goat serum for 30 min at room temperature and were incubated with primary antibodies to IBA1 (1:500, 019-19741, Wako) and GFAP (1:500, G3893, Sigma–Aldrich) at 4 °C for 12 h. The sections were then incubated with fluorescent-labeled goat anti-rabbit (1:500, 111-545-144, Jackson ImmunoResearch) and goat anti-mouse (1:500, 115-165-075, Jackson ImmunoResearch). The related fluorescence intensities were analyzed by image pro plus software.

Wang B., Ma Y., Li S., Yao H., Gu M., Liu Y., Xue Y., Ding J., Ma C., Yang S, & Hu G. (2023). GSDMD in peripheral myeloid cells regulates microglial immune training and neuroinflammation in Parkinson's disease. Acta Pharmaceutica Sinica. B, 13(6), 2663-2679.

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Quantifying Dopaminergic Neurons in Zebrafish

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The immunostaining and counting of TH+ neurons in zebrafish were performed as previously described^{42 (link)}. Briefly, adult fish were sacrificed by the addition of 0.1% tricaine to the circulating water of the breeding system. Brains were removed, explanted, and fixed with 4% (w/v) paraformaldehyde in PBS (Nacalai) at 4 °C O/N. The specimens were embedded in 2% low-melting agarose, and 200 μm axial sections were prepared using a PRO7 microslicer (Dosaka EM, Kyoto, Japan). Floating slices were incubated with 10 mM sodium citrate buffer, pH 8.5, at 80 °C for 120 min. After washing with PBS containing 1% Triton X-100, sections were blocked with 2% BSA in PBS/1% Triton X-100 buffer for 30 min. These pretreated sections were incubated with a rabbit anti-TH antibody (1/500, AB152, EMD-Millipore, Billerica, MA, USA) O/N at 4 °C. After washing with PBS/1% Triton X-100 buffer, the sections were incubated with anti-rabbit IgG conjugated to Alexa Fluor 594 (1/100, ThermoFisher Scientific) O/N at 4 °C. The sections were washed with PBS/1% Triton X-100 buffer and analyzed using an A1R+ confocal microscope (Nikon).

Matsui H., Ito J., Matsui N., Uechi T., Onodera O, & Kakita A. (2021). Cytosolic dsDNA of mitochondrial origin induces cytotoxicity and neurodegeneration in cellular and zebrafish models of Parkinson’s disease. Nature Communications, 12, 3101.

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Quantifying CrebB Expression in Drosophila Brains

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Flies, in which CrebB knockout was induced, were moved for 6 days to 29°C before antibody staining experiments. Male adult brains were dissected in phosphate-buffered-saline (PBS) and fixed at room temperature for 30 min with a 3.7% formaldehyde solution (in PBS). Brains were washed at least five times with PBST (PBS with 0.3% triton X-100) before primary antibodies were added for overnight incubation at 4°C. The following primary antibodies were used: rabbit anti-Eyeless (1:400, courtesy of Uwe Waldorf), chicken anti-GFP (1:1000, Abcam ab13970), rabbit anti-GFP (1:1000, Invitrogen A-6455), guinea pig anti-CrebB (1:400), rabbit anti-Tyrosine hydroxylase (1:100, Merck AB152), mouse anti-Repo (1: 20, Developmental Studies Hybridoma Bank 8D12), mouse anti-Discs large (1:50, Developmental Studies Hybridoma Bank 4F3). Brains were washed again before the overnight incubation at 4°C with secondary antibodies. The used secondary antibodies were conjugated with Alexa fluorescent proteins (488, 567 or 647; Molecular Probes) and diluted 1:200. After a last washing step, brains were mounted in Vectashield H-1000 or Vectashield with DAPI H-1200 (Vector Laboratories) on a microscope slide. Samples were imaged with Leica SP5 confocal microscope and the images were processed with Imaris Bitplane 9.2 and Adobe Photoshop CS6.

Widmer Y.F., Fritsch C., Jungo M.M., Almeida S., Egger B, & Sprecher S.G. (2018). Multiple neurons encode CrebB dependent appetitive long-term memory in the mushroom body circuit. eLife, 7, e39196.

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Immunoblotting of Dopaminergic Markers

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This was performed as previously described [20 (link)] with antibodies against D2R (1:500, rabbit), dopamine transporter (DAT; 1:500, rabbit), tyrosine hydroxylase (TH; 1:1000, rabbit) (AB5084P, AB1591P and AB152, Merck Millipore, Billerica, MA, USA), signal transducer and activator of transcription 3α (STAT3α; 1:1000, rabbit), DNMT1 (1:1000, rabbit), DNMT3a (1:1000, rabbit) (nos. 8768, 5032 and 3598; Cell Signaling Technology, Tokyo, Japan), DNMT3b (1 μg/ml, rabbit), ERRγ (1:1000, rabbit) and β-actin (1:10,000, mouse) (ab16049, ab128930 and ab6276; Abcam, Cambridge, MA, USA).

Kozuka C., Kaname T., Shimizu-Okabe C., Takayama C., Tsutsui M., Matsushita M., Abe K, & Masuzaki H. (2017). Impact of brown rice-specific γ-oryzanol on epigenetic modulation of dopamine D2 receptors in brain striatum in high-fat-diet-induced obesity in mice. Diabetologia, 60(8), 1502-1511.

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Immunohistochemical Staining of TH and TPH2

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A polyclonal antibody to TH (catalog No. AB152, EMD Millipore Corporation, Billerica, MA, USA) and TPH2 (catalog No. ABN60, EMD Millipore Corporation) were used for immunohistochemistry as the primary antibody. Fluorescein isothiocyanate-labeled goat anti-rabbit IgG (catalog No. 65-6111, Invitrogen) was used as the secondary antibody. The immunohistochemical staining procedure was performed according to previously reported methods and conditions (Sutoo et al. 2002 (link)). Slices were rinsed with PBS four times for a total of 60 min. Primary antibodies were diluted 1:200 for TH and 1:400 for TPH2 with PBS and placed on each brain slice, then reacted for 12 h at 4 °C in a moist dark box. After reaction, the slices were rinsed four times with PBS for a total of 60 min. The secondary antibody was diluted 1:100 with PBS, placed on each slice, and incubated at 24 °C for 3 h in a moist dark box. After the reaction, the slices were rinsed four times with PBS for a total of 60 min. Finally, these slides were embedded in 10% glycerin in PBS and kept moist at 4 °C in a dark box.

Nasu M., Yada S., Igarashi A., Sutoo D., Akiyama K., Ito M., Yoshida N, & Ueda S. (2014). Mammalian-Specific Sequences in Pou3f2 Contribute to Maternal Behavior. Genome Biology and Evolution, 6(5), 1145-1156.

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Multimodal Analysis of Mouse Brain

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Mouse brain tissue was prepared by fixation via transcardial perfusion with 4% paraformaldehyde. Brains were then removed and equilibrated in graded sucrose solutions, frozen at −8°C in OCT solution, and cryosectioned at 25 μm for fluorescence/immunofluorescence imaging. Tissue processed in this way was suitable for imaging of endogenous protein fluorescence, immunofluorescence, and fluorescence in situ hybridization (FISH). Tyrosine hydroxylase (TH) immunoreactivity was detected using rabbit anti-TH (AB152, EMD Millipore, RRID:AB_390204). Multi-channel FISH was performed with the RNAscope Multiplex Fluorescent V2 kit, according to the manufacturer’s instructions (Advanced Cell Diagnostics). The probes used included: EGFP, #400281-C2 (channel 2) and Mm-Rln3-C1 (channel 1).

Nasirova N., Quina L.A., Novik S, & Turner E.E. (2021). Genetically Targeted Connectivity Tracing Excludes Dopaminergic Inputs to the Interpeduncular Nucleus from the Ventral Tegmentum and Substantia Nigra. eNeuro, 8(3), ENEURO.0127-21.2021.

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Western Blot Analysis of Neuronal Targets

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The protocol for Western blot was performed as previously described61 (link). Primary antibodies (anti α1 1:2000 (a6f-c, DevelopmenalStudies Hybridoma Bank), anti α2 1:1000 (07674, EMD Millipore, US), anti α3 1:1000 (06172, EMD Millipore, US), anti DAT 1:500 (MAB369, EMD Millipore, US), anti TH 1:2000 (AB152, EMD Millipore, US) and GAPDH 1:1000 (ab9485, Abcam, Cambridge, UK)) were incubated overnight at 4 °C. Next day, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (swine anti-rabbit 1:2000 (Dako, Glostrup, Denmark), rabbit anti-mouse 1:2000 (Dako, Glostrup, Denmark)) for 1 hour at room temperature. Visualization was done with a LAS 3000 imager (Fujifilm, Tokyo, Japan) using Amersham ECL Western Blotting Detection Kit (GE Healthcare, Buckinghamshire, UK) as the detection reagent. ImageJ version 1.48 v was used for densitometric analysis of the Western blots.

Holm T.H., Isaksen T.J., Glerup S., Heuck A., Bøttger P., Füchtbauer E.M., Nedergaard S., Nyengaard J.R., Andreasen M., Nissen P, & Lykke-Hartmann K. (2016). Cognitive deficits caused by a disease-mutation in the α3 Na+/K+-ATPase isoform. Scientific Reports, 6, 31972.

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Oxidative Stress Immunohistochemistry Protocol

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The free-floating sections were immune-blocked with 4% goat serum in 0.25% triton/PBS for 2 hours and then incubated with 3-Nitrotyrosine (3-NT, mouse, 1:200, Abcam, Cambridge, MA), 4-Hydroxynonenal (4-HNE, rabbit, 1:200, Abcam, Cambridge, MA) or gp91^phox antibody (mouse, 1:500, BD Biosciences, 611414) antibodies overnight at 4 °C. On the second day, the sections were washed with 1% BSA in 0.25% triton/PBS before incubation with polyclonal rabbit anti-TH antibody (mouse: MAB318, 1:2000; rabbit: AB152, 1:2000; EMD Millipore, Temecula, CA) overnight at 4 °C. The double-label immunofluorescence pictures were taken under the confocal microscope using Alexa-488 (green) and Alexa-594 (red) conjugated secondary antibodies (1:1,000) to visualize the double TH and 3-NT, 4-HNE or gp91^phox-positive neurons. Densitometry was performed using ImageJ.

Song S., Wang Q., Jiang L., Oyarzabal E., Riddick N.V., Wilson B., Moy S.S., Shih Y.Y, & Hong J.S. (2019). Noradrenergic dysfunction accelerates LPS-elicited inflammation-related ascending sequential neurodegeneration and deficits in non-motor/motor functions. Brain, behavior, and immunity, 81, 374-387.

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Transcardial Perfusion and Brain Histology

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Mice were euthanized using fatal plus at the desired time points after injection. Under deep anesthesia, mice were rapidly transcardially perfused with 15ml of ice-cold 1X PBS containing heparin for approximately 2 miniutes, followed by perfusion with ice-cold 4% paraformaldehyde (PFA; 25 ml at 5 ml/min). Brains were further post-fixed in 4% PFA for 24 hours, then transferred to ice-cold 10% sucrose solution and incubated at 4°C overnight. The 10% sucrose were then replaced with ice-cold 30% sucrose, and brains were allowed to equilibrate completely at 4 °C, as evidenced by their sinking to the bottom of the container. Brains were then sectioned immediately on a freezing sliding microtome and processed for immune-staining as described previously (Wang et al., 2014 (link); Wang et al., 2015 (link)). We used the following primary antibodies for immunohistochemistry: antibodies against tyrosine hydroxylase (TH, rabbit: AB152, 1:4000; EMD Millipore, Temecula, CA), neuronal nuclei (Neu-N, 1:4000, CHEMICON, Temecula, CA), ionized calcium binding adaptor molecule-1 (Iba-1; 1:5000, Wako Chemicals, Richmond, VA), and CD11b (1:400, AbD Serotec, Raleigh, NC). Immuno-staining was visualized by using 3,3′-diaminobenzidine (DAB) and urea-hydrogen peroxide tablets or nickel-enhanced DAB.

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