Map2 antibody

Manufactured by Abcam

Sourced in United States, United Kingdom

The MAP2 antibody is a reagent used in research applications to detect the presence and distribution of the microtubule-associated protein 2 (MAP2) in biological samples. MAP2 is a structural protein found primarily in the dendrites of neurons and plays a crucial role in the organization and stability of the neuronal cytoskeleton. This antibody can be used in techniques such as immunohistochemistry, immunocytochemistry, and Western blotting to visualize and quantify MAP2 expression in various cell and tissue types.

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Lab products found in correlation

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Spelling variants of this product

Anti-MAP2 antibody (15 citations) Mouse anti-MAP2 antibody (3 citations) Anti-MAP2 antibody (ab5392) (2 citations) Rabbit anti-MAP2 monoclonal antibody (2 citations)

13 protocols using map2 antibody

5-HT6 Receptor Mutagenesis and Localization

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The mouse EGFP-tagged 5-HT6 plasmid was provided by Kirk Mykytyn (Ohio State University, USA). We constructed 13 mutagenesis plasmids. In each clone, the codons for amino acids 69, 70, 72, 106, 230, 234, 262, 265, 284, and 350 were replaced by triplet nucleotides. Mouse GFP-tagged ARL13B was amplified from cDNA from the mouse brain. 5-HT6-mcherry plasmid was constructed. PEGFP-N3 and mcherry-N3 were purchased from Addgene.
SB271046 drug (Santa Cruz Biotechnology), 5-HT6R siRNA (QIAGEN), siRNAs of 5-HT1A, 5-HT1B, 5-HT1D, 5-HT1F, 5-HT2A, 5-HT2B, 5-HT2C, 5-HT3A, 5-HT4, 5-HT5A, 5-HT5B, and 5-HT7 (Santa Cruz Biotechnology), 5-HT6R antibody (Abcam), MAP2 antibody (Abcam), AC3 antibody (Santa Cruz Biotechnology), SSTR3 antibody (Santa Cruz Biotechnology), ARL13B antibody (NeuroMab), ankyrin G antibody (Invitrogen), neurofascin 186 antibody (Abcam), γ-tubulin antibody (Sigma), IFT88 antibody (Proteintech), calbindin antibody (Swant), CamkII antibody (Millipore), Gad antibody (Abcam), Nav1.1 antibody (NeuroMab), Nav1.6 antibody (Alomone), Nav1.2 antibody (NeuroMab), Kv1.1 antibody (Alomone), and AlexaFluor fluorescence antibodies (Invitrogen) were utilized.

Hu L., Wang B, & Zhang Y. (2017). Serotonin 5-HT6 receptors affect cognition in a mouse model of Alzheimer’s disease by regulating cilia function. Alzheimer's Research & Therapy, 9, 76.

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Immunofluorescence analysis of α-synuclein PFFs in primary cortical neurons

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Primary cortical neurons were dissected from the cortex of embryonic day (E) 16–E18 Sprague Dawley rats (Shanghai SIPPR BK Laboratory Animals Ltd, China) embryos as previously described^{41 (link)}. In brief, primary neurons were seeded onto coverslips previously coated with poly-D-lysine (PDL) in 24-well plates (150,000 cells/coverslip). At 8-day in vitro (DIV), neurons were treated with PBS and 100 nM (final concentration) WT_1a or G51D α-syn PFFs, and collected for immunofluorescence at 10/14-day post-treatment. The primary antibodies used in the assay were the phospho-α-synuclein (S129) antibody (Abcam, cat. no. ab51253) and the MAP2 antibody (Abcam, cat. no. ab5392). The antibodies were diluted at a ratio of 1:1000. The intensity of confocal images was analyzed by Image J 2.0.0. The secondary antibodies included goat anti-rabbit IgG Alexa Fluor 568 (Abcam, cat. no. ab175471) and goat anti-chicken Alexa Fluor 488 (Thermo Fisher, cat. no. A-11039). All rat experiments were performed followed the protocols approved by the Animal Care Committee of the Interdisciplinary Research Center on Biology and Chemistry (IRCBC), Chinese Academy of Sciences (CAS). There are three samples in each group.

Sun Y., Long H., Xia W., Wang K., Zhang X., Sun B., Cao Q., Zhang Y., Dai B., Li D, & Liu C. (2021). The hereditary mutation G51D unlocks a distinct fibril strain transmissible to wild-type α-synuclein. Nature Communications, 12, 6252.

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Immunocytochemical Analysis of Cell Cultures

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Cells were cultured on poly-D-lysine coated coverslips, fixed with 4% paraformaldehyde/PBS for 10 min and permeabilised with 0.2% Triton X100/PBS for 20 min. Blocking was carried out with 10% BSA in 0.1% Triton X100/PBS for 30 min. Primary antibodies (MYC antibody (9E10), Sox2 antibody (R&D, MAB2018), MAP-2 antibody (Abcam)) and the fluorescent-conjugated secondary antibodies were incubated at room temperature for 1 h. Staining was observed after mounting in mounting medium for fluorescence with DAPI (Vector). Error bars in Figures
1C based on counting 15 cells and
3A, and
4A counting 100 cells.

Liu Y.R., Laghari Z.A., Novoa C.A., Hughes J., Webster J.R., Goodwin P.E., Wheatley S.P, & Scotting P.J. (2014). Sox2 acts as a transcriptional repressor in neural stem cells. BMC Neuroscience, 15, 95.

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Molecular Analysis of Alzheimer's Pathology

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Human Aβ_1–42 was purchased from EMD Millipore Corporation (Billerica, MA, USA). Ori was obtained from Chengdu Must Bio-Technology Company (Sichuan, China). Neurobasal media and B27 were bought from Life Technologies Corporation. MAP-2 antibody was purchased from Abcam (Cambridge, MA, USA). COX-2 antibody was bought from Santa Cruz Biotechnology (Santa Cruz, USA). PSD-95 antibody, synaptophysin antibody and CREB antibody were obtained from CST (Cell Signaling Technology, USA). 4', 6'-diamidino-2-phenylindole (DAPI), VDAC1, BDNF, p-TrkB, TrkB, p-CREB and β-actin antibodies were purchased from Bioworld Technology (Bioworld, USA). Lamin B1 antibody and HRP-conjugated secondary antibodies were also obtained from Bioworld.

Wang S., Yu L., Yang H., Li C., Hui Z., Xu Y, & Zhu X. (2016). Oridonin Attenuates Synaptic Loss and Cognitive Deficits in an Aβ1–42-Induced Mouse Model of Alzheimer’s Disease. PLoS ONE, 11(3), e0151397.

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Neuronal Cell Culture Protocols

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Neurobasal Medium, Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), Hank’s balanced salt solution (HBSS), B-27 Serum-Free Supplement, L-glutamine and 0.25% trypsin-EDTA were purchased from Gibco, Langley, OK, USA. Cell culture flasks, 6-well plates, 96-well plates and Pasteur pipettes were purchased from Corning Inc., Corning, NY, USA. Deuterium oxide (D₂O), TMSP-2,2,3,3-D4 (D, 98%) and sodium-3-trimethylsilylpropionate (TSP) were purchased from Cambridge Isotope Laboratories, Tewksbury, MA, USA. D-(+)-glucose, streptozotocin (STZ) and ATP, ADP, AMP standards were purchased from Sigma Aldrich, St. Louis, MO, USA. Poly-D-lysine and the penicillin-streptomycin solution were purchased from Beyotime Biotechnology, China. The MAP2 antibody and goat anti-rabbit IgG-FITC antibody were obtained from Abcam, Britain and Santa Cruz Biotechnology, Santa Cruz, CA, USA, respectively.

Zhao L., Dong M., Wang D., Ren M., Zheng Y., Zheng H., Li C, & Gao H. (2018). Characteristic Metabolic Alterations Identified in Primary Neurons Under High Glucose Exposure. Frontiers in Cellular Neuroscience, 12, 207.

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Axonal Length Quantification in Neurons

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After treatment with the CM, recombinant pyruvate kinase isoform M2 (PKM2; 0.01, 0.1, 1, and 10 ng/mL), or vehicle solution, neurons were fixed with paraformaldehyde for 90 min. Axons were immunostained with the mouse monoclonal anti-phosphorylated neurofilament-H (pNF-H) antibody (1:250; Covance Inc., Princeton, NJ). Neuronal cell bodies were immunostained with the rabbit polyclonal anti–microtubule-associated protein 2 (MAP2) antibody (1:2000; Abcam, Cambridge, UK). As secondary antibodies, we used Alexa Fluor-488-conjugated goat anti-mouse immunoglobulin G (IgG) and Alexa Fluor-568-conjugated goat anti-rabbit IgG (1:300; Thermo Fisher Scientific). Nuclear staining with 1 μg/mL 4′6-diamino-2-phenylindole (DAPI) was performed at room temperature. Eight to 23 images per well were captured using a fluorescent microscope (Axio Cell Observer Z1; Carl Zeiss, Oberkochen, Germany), at a field of 432 μm × 323 μm or 863 μm × 645 μm. The length of pNF-H–positive axons was measured using MetaMorph software (Molecular devices, San Jose, CA), which automatically traces and measures neurite lengths. The total axonal length in a given image was divided by the number of MAP2- and DAPI-double positive cells to calculate the axonal length per neuron.

Kodani A., Kikuchi T, & Tohda C. (2019). Acteoside Improves Muscle Atrophy and Motor Function by Inducing New Myokine Secretion in Chronic Spinal Cord Injury. Journal of Neurotrauma, 36(12), 1935-1948.

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Immunostaining of Cortical Neurons

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Cortical neurons were fixed with paraformaldehyde for 15 min, and then permeabilized with 0.3% Triton X-100 for 30 min. After washing with PBS, the cells were blocked with 5% BSA for 1 h at room temperature, followed by incubation with a MAP2 antibody (1:100; Abcam, UK), a NeuN antibody (1:100; Millipore, USA), and an SH3RF2 antibody (1:100; Novus, USA) overnight at 4°C. The day after, neurons were incubated with Alexa Fluor 488-conjugated (1:500; Abcam, UK) and Alexa Fluor 555-conjugated (1:500; Abcam, UK) for 1 h at room temperature. Finally, nuclei were visualized with DAPI (1:1,000; Abcam, UK). Images were acquired with a confocal laser scanning microscope (LSM 780; Zeiss, Germany) and analyzed by Image J software.

Chen J.Q., Gao S.Q., Luo L., Jiang Z.Y., Liang C.F., He H.Y, & Guo Y. (2022). Nonoxid-HMGB1 Attenuates Cognitive Impairment After Traumatic Brain Injury in Rats. Frontiers in Medicine, 9, 827585.

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Isolation and Culture of Primary Neurons

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The separation of primary neurons of neonatal rats was performed as described previously [18] (link). The separated primary neurons were adjusted to about 1×10⁶ cells/ml with 1 ml inoculated into each well of a 6-well plate precoated with poly-D-lysine (50 µg/ml; Sigma, U.S.). The culture medium was changed every 48 h. The purity of neurons was over 95% after 8 days, as examined by Map2 antibody (Abcam, U.K.). The primary neurons were used after 8 days of culturing [24] (link).

Chai Y.S., Yuan Z.Y., Lei F., Wang Y.G., Hu J., Du F., Lu X., Jiang J.F., Xing D.M, & Du L.J. (2014). Inhibition of Retinoblastoma mRNA Degradation through Poly (A) Involved in the Neuroprotective Effect of Berberine against Cerebral Ischemia. PLoS ONE, 9(3), e90850.

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Antibody and Reagent Selection Guide

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The following antibodies and reagents were used: VCP antibody was purchased from Cell signaling Technology (Danvers, MA, USA) or Abcam (Cambridge, United Kingdom). β-actin, calpain2, GSK-3β (S9), mTOR (S2448) and cleaved caspase-3 antibodies were purchased from Cell Signaling Technology; LC3B antibody from Sigma-Aldrich (St. Louis, MO, USA); MAP2 antibody from Abcam; GFAP and Vimentin antibodies from Millipore (Billerica, MA, USA). DBeQ, staurosporine, lactacystin and BafA1 were purchased from Sigma-Aldrich, Z-VAD-FMK from R&D Systems (Minneapolis, MN, USA), necrostatin-1 from Enzo Life Sciences (Farmingdale, NY, USA). They were diluted in dimethyl sulfoxide at appropriate concentrations.

Yeo B.K., Hong C.J., Chung K.M., Woo H., Kim K., Jung S., Kim E.K, & Yu S.W. (2016). Valosin-containing protein is a key mediator between autophagic cell death and apoptosis in adult hippocampal neural stem cells following insulin withdrawal. Molecular Brain, 9, 31.

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Neuronal Cell Culture Protocol

Cited 1 time

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Hanks' balanced salt solution (HBSS), serum-free neurobasal media (NB), B27 supplement (50x), L-glutamine, Prolong Gold anti-fade reagent with 4′,6-diamidino-2-phenylindole (DAPI), SuperScript III reverse transcriptase, SYBR/Green Master mix, and Alexa 405-, 488-, or 555-conjugated secondary antibodies were obtained from ThermoFisher. Poly-L-lysine hydrobromide, cytosine β-D-arabinofuranoside hydrochloride (Ara-C), and anti-actin antibody were from Sigma. Microtubule-associated protein 2 (MAP-2) antibody was obtained from Abcam. C5aR1 antibody (10/92) was obtained from SeroTec (Ager et al., 2010 (link)). Anti VGLUT1 and anti GAD67 antibodies were obtained from Millipore. HRP-conjugated F(ab')₂ antibodies were from Jackson ImmunoResearch Laboratories. Human C5a peptide was obtained from CompTech. PMX53 was obtained from Cephalon. All buffers are made with Millipore-purified water with additional filters to eliminate LPS and are periodically tested for endotoxin using the Limulus amebocyte lysate clot assay (all solutions added to cells were <0.1 EU/mL; 1 EU is equivalent to 0.1 ng/mL LPS).

Hernandez M.X., Namiranian P., Nguyen E., Fonseca M.I, & Tenner A.J. (2017). C5a Increases the Injury to Primary Neurons Elicited by Fibrillar Amyloid Beta. ASN NEURO, 9(1), 1759091416687871.

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