The purified polyclonal antibody against SNPH residues 225–428 was described previously (Kang et al., 2008 (link)). Sources of other antibodies or reagents are as follows: anti–βIII-tubulin, anti-MAP2, anti–Tau-1, anti-dynein IC74, and anti–GAP-43 were from EMD Millipore; anti–cytochrome c was from BD; anti-TOM20 was from Santa Cruz Biotechnology, Inc.; anti-Miro1/2 (HPA010687) and anti-Trak2 were from Sigma-Aldrich; ECL-HRP–linked secondary antibodies were from GE Healthcare; and Alexa Fluor 546– or Alexa Fluor 488–conjugated secondary antibodies were from Invitrogen.
Anti map2
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany
Anti-MAP2 is a laboratory reagent used for the detection and quantification of microtubule-associated protein 2 (MAP2) in various biological samples. MAP2 is a cytoskeletal protein that is primarily found in the dendrites of neurons and plays a crucial role in the stabilization and organization of the neuronal cytoskeleton. The Anti-MAP2 product can be used in techniques such as immunohistochemistry, Western blotting, and enzyme-linked immunosorbent assays (ELISA) to study the expression and distribution of MAP2 in research applications.
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Lab products found in correlation
Anti gfap, by Merck Group (19 mentions)
Anti neun, by Merck Group (13 mentions)
Anti nestin, by Merck Group (12 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (11 mentions)
Anti tuj1, by Merck Group (10 mentions)
Anti gfap, by Agilent Technologies (7 mentions)
Anti sox2, by Santa Cruz Biotechnology (6 mentions)
Anti flag, by Merck Group (6 mentions)
Lsm 710 confocal microscope, by Zeiss (6 mentions)
Anti sox2, by Merck Group (6 mentions)
Anti ctip2, by Abcam (6 mentions)
Anti sox2, by Abcam (5 mentions)
Alexa 488, by Thermo Fisher Scientific (5 mentions)
Anti psd95, by Thermo Fisher Scientific (5 mentions)
Anti lc3b, by Cell Signaling Technology (4 mentions)
Anti nanog, by Abcam (4 mentions)
Anti pax6, by Fortrea (4 mentions)
Anti actin, by Merck Group (4 mentions)
Lsm 700, by Zeiss (4 mentions)
Anti gfp, by Abcam (4 mentions)
111 protocols using anti map2
1
Immunofluorescence Staining Antibody Protocol
2
Antibody Characterization for Cellular Pathways
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The following antibodies were used in this study: anti-SARA [mouse, sc-133071; 1:200 for immunoblotting and 1:100 for immunofluorescence (IF)], anti-PP1c (mouse, sc-7482; 1:50 for IF), anti-GADD34 (mouse, sc-373815; 1:50 for IF), anti-TβRI (mouse, sc-101574; 1:100 for IF) and anti-Smad2/3 (mouse, sc-398844; 1:100 for IF); all these antibodies were purchased from Santa Cruz Biotechnology (Dallas, Texas, United States). The antibody anti-MAP2 (rabbit, 1:500) and anti-Tau-1 (mouse, 1:500) were from Merck Millipore (Darmstadt, Germany). Antibody anti-pSamd2/3 (rabbit, s465/s467, E8F3R, 1:50 for IF) was from Cell Signaling (Danvers, MA, United States) and anti-phospho TβRI (rabbit, ser165, Lot: DY1241; 1:50 for IF) was acquired from Elabscience (Houston, Texas, United Sates).
3
Immunohistochemical Characterization of Cells
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Anti-ACE2 (Santa Cruz Biotechnology, SC-390851, Dallas, TX, USA), anti-SATB2 (Abcam, ab51502, Eugene, OR, USA), anti-beta III Tubulin (TUJ-1) (Abcam, ab14545), anti-MAP2 (EMD Millipore, AB2290, Burlington, MA, USA), Anti-LaminA/C (Cell Signaling Technology, 2032, Danvers, MA, USA), and anti-SOX2 (Abcam, ab59776) antibodies were used as primary antibodies for immunohistochemistry, immunocytochemistry, and immunoblot assays. Goat anti-Rabbit IgG (H+L), TRITC (Thermo Fisher Scientific, A16101, Waltham, MA, USA), and goat anti-Mouse IgG (H+L), Alexa Fluor 488 (Thermo Fisher Scientific, A11001) were used as secondary antibodies for immunohistochemistry and immunocytochemistry. Goat anti-Rabbit IgG (H+L), HRP (Thermo Fisher Scientific, 31460) and goat anti-Mouse IgG (H+L), HRP (Thermo Fisher Scientific, 31430) were used as secondary antibodies for immunoblot assay.
4
Immunofluorescence Analysis of Neuronal Markers
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For immunofluorescence analysis, DIV14-21 neurons in the microfluidic device were fixed with 4% paraformaldehyde (PFA) for 30 min at room temperature (RT). The fixation buffer was rinsed out with PBS, after which neurons were permeabilized with 0.2% Triton X-100 for 30 min and subsequently incubated for 30 min with a blocking solution containing 5% bovine serum albumin (BSA). Anti-Tau (Merck Millipore), anti-MAP2 (Merck Millipore), anti-synapsin (Synaptic Systems), and anti-Shank3 (Synaptic Systems) antibodies were applied to microchannels and the microfluidic device was incubated overnight at 4 °C. After incubating with primary antibodies, neurons were washed with PBS for 30 min and subsequently incubated with Alexa 488- and/or Alexa 546-conjugated secondary antibodies (Invitrogen) for 1 h, providing different color combinations as needed.
5
Proteomic analysis of Alzheimer's markers
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The following primary antibodies were used in this study: anti-Aβ (82E1, IBL, Gunma, Japan; 6E10,Covance, Emeryville, CA, USA), anti-BACE1 (AB5832, Merck Millipore, Darmstadt, Germany; D10E5, Cell Signaling, Danvers, MA, USA), anti-APP (R37 [50 (link)]; 22C11, Merck Millipore), anti-ADAM10 (Sigma, St Louis, MO, USA), anti-phospho-eIF2α (Ser51) (Cell Signaling), anti-eIF2α (Assay Biotechnology, Sunnyvale, CA, USA), anti-cleaved caspase 3 (Asp175) (Cell Signaling), anti-GRP78 (BD Biosciences, San Jose, California, USA), anti-β-actin (Sigma), anti-MAP2 (Merck Millipore), and antibody to the rhodopsin tag (1D4) [51 (link)] obtained from University of British Columbia.
6
Immunofluorescence and Western Blot Antibody Protocol
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In immunofluorescence experiments, the following antibodies and dilutions were used: anti-Phospho SQSTM1/p62 (S349) (Abcam, Cambridge, UK, cat # ab211324) 1:100; anti-SQSTM1/p62 (Abcam, cat # ab56416) 1:50; anti-β-Tubulin III (Merck Millipore, Burlington, MA, USA cat # T2200) 1:500; anti-MAP2 (1:500, Merck Millipore, cat # M9942) 1:500; anti-GFAP (Thermo Fisher Scientific, Waltham, MA, USA, cat # 13-0300), 1:800, donkey anti-rabbit-IgG Alexa Fluor 488 (Thermo Fisher Scientific, cat # R37118) 1:1000; donkey anti-mouse-IgG Alexa Fluor 594 (Thermo Fisher Scientific, cat # A-21203, 1:1000); goat anti-mouse IgG1 CF 568 (Merck, cat # SAB4600313 1:1000); goat anti-rat Alexa Fluor 647 (Thermo Fisher Scientific, cat # A21247, 1:1000); and DAPI (4′,6-diamidino-2-phenylindole Merck Millipore, cat #D9542) 1:5000. In Western blotting experiments, the following antibodies and dilutions were used: anti-Phospho SQSTM1/p62 (S349) (Abcam cat # ab211324) 1:2000; anti-SQSTM1/p62 (Abcam cat # ab56416) 1:2000; Anti-GAPDH (Abcam cat # ab8245); 1:5000; anti-phospho-Akt (Ser473 D9E Cell Signaling, Danvers, MA, USA, cat #4060) 1:2000; anti-Akt (Cell Signaling, cat # 9272, 1:1000); goat anti-mouse IgG IRDye 800(Li-Cor, Lincoln, NE, USA, cat # 926-32210) 1:5000; and goat anti-rabbit-IgG Alexa 680 (Thermo Fisher Scientific cat # A21076) 1:5000.
7
Comprehensive Immunofluorescence Staining of Brain Regions
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All brain tissue processed for immunofluorescence (IF) was frozen with optimal cutting temperature compound on a microtome stage and sectioned/collected for ST (25 μm in thickness) and SN (40 μm in thickness) regions. Tissue sections were stored in cryoprotectant (30% sucrose, 30% ethylene glycol, and 0.5 M phosphate buffer, pH 7.2) at 20°C until selected for immunostaining. IF staining was conducted as previously described by Miller et al. (2011) (link), and all antibody dilutions were 1:500, unless stated otherwise (Miller et al., 2011 (link)). For stereology, SN sections were immunostained with anti-TH (cat. no. AB152; Merck Millipore, Burlington, MA) and anti-MAP2 (cat. no. AB5392; Abcam). For gliosis, SN and ST sections were immunostained with either anti–ionized calcium-binding adapter molecule 1 (IBA-1) (1:250; cat. no. 016-20001; Wako Chemicals USA, Inc., Richmond, VA) or anti–glial fibrillary acidic protein (GFAP) (cat. no. Z0334; Agilent Technologies, Santa Clara, CA) and anti-TH (cat. no. AB76442, Abcam). SN tissue was also immunostained with anti-TH and anti-Nurr1 (1:200, SC991; Santa Cruz Biotechnology, Dallas, TX) for mean intensity measurements of Nurr1. All secondary antibodies used for IF were Alexa Fluor 488, 555, and 647 (Thermo Fisher Scientific, Carlsbad, CA).
8
Western Blot Analysis of Microglia and Neurons
9
Evaluating Neuritic Outgrowth and Synaptic Connectivity
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To evaluate neuritic outgrowth and connectivity, fixed cells were stained with the following primary antibodies: a rabbit polyclonal anti-MAP2 (1:100, Merck Millipore, Burlington, MA, USA) and a mouse monoclonal anti-PSD-95 (1:100, Abcam, Cambridge, UK) antibody. Detection was performed using a FITC-conjugated anti-rabbit IgG (1:200, Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) and a Cy3-conjugated anti-mouse IgG (1:200, Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) antibody, respectively. Nuclei were counterstained with Hoechst-33342 (Sigma-Aldrich, St. Louis, MO, USA) and fluorescent images were acquired using a Nikon Eclipse Te600 microscope equipped with a Nikon Digital Camera DXM1200 ATI system (Nikon Instruments, Inc., Melville, NY, USA). The neuritic length of MAP2-positive neurons was measured and quantified by tracing along each neuronal projection using the image analysis system Image Pro Plus (Media Cybernetics, Silver Spring, MD, USA) as previously described [11 (link)]. The degree of synaptic innervation was evaluated by counting the number of PSD-95 positive puncta on proximal dendrites and expressed as the number of PSD-95 puncta per 20 μm of neuritic length. Immunofluorescence images were taken with a LEICA TCS SL confocal microscope (Leica Mycrosystems, Wetzlar, Germany). Fifty neurons for each condition were evaluated.
10
Immunoblotting and Immunofluorescence Analysis
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We used anti-FMRP (Abcam; Ref. ab69815), anti-MAP2 (Merck Millipore; Ref.
MAB378), and anti-LC3B (Cell Signaling; Ref. #2775) as primary antibodies, while horseradish peroxidase (HRP)-conjugated anti-rabbit (Promega), Alexa Fluor 555 goat anti-rabbit (Invitrogen), and Alexa Fluor 488 goat anti-mouse (Invitrogen) were used as secondary antibodies. homogenization in extraction buffer containing SDS and protease and phosphatase inhibitor cocktails. Protein extracts were resolved by SDS-PAGE on Mini-PROTEAN® TGX Stain-Free™ precast gels (BioRad). Proteins were transferred to polyvinylidene difluoride (PVDF) membranes. HRP-conjugated secondary antibodies were detected with the WesternBright Quantum HRP Substrate (Advansta) and chemiluminiscence measured on a ChemiDoc XRS (BioRad) imager.
MAB378), and anti-LC3B (Cell Signaling; Ref. #2775) as primary antibodies, while horseradish peroxidase (HRP)-conjugated anti-rabbit (Promega), Alexa Fluor 555 goat anti-rabbit (Invitrogen), and Alexa Fluor 488 goat anti-mouse (Invitrogen) were used as secondary antibodies. homogenization in extraction buffer containing SDS and protease and phosphatase inhibitor cocktails. Protein extracts were resolved by SDS-PAGE on Mini-PROTEAN® TGX Stain-Free™ precast gels (BioRad). Proteins were transferred to polyvinylidene difluoride (PVDF) membranes. HRP-conjugated secondary antibodies were detected with the WesternBright Quantum HRP Substrate (Advansta) and chemiluminiscence measured on a ChemiDoc XRS (BioRad) imager.
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