13-1500

antibody from Invitrogen Antibodies

Targeting: MAP2 MAP2A, MAP2B, MAP2C

Provider product page for 13-1500

Western blot

ELISA

Immunoprecipitation

Antibody data

Antibody Data
Antigen structure
References [52]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [2]
Immunohistochemistry [1]
Flow cytometry [1]
Other assay [22]

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Product number: 13-1500 - Provider product page
Provider: Invitrogen Antibodies
Product name: MAP2 Monoclonal Antibody (M13)
Antibody type: Monoclonal
Antigen: Other
Description: This antibody reacts with a 300 kD cytoskeletal protein (MAP2A and MAP2B) primarily expressed in the somatodendritic domain of the neuron but is also found in the glia. Reactivity is not affected by phospohorylation state of MAP2. Abnormal expression of MAP2 may cause cytoskeletal abnormalities and impair signal transduction in dendrites. This antibody is suitable for immunohistochemical staining of alcohol-fixed paraffin embedded or frozen tissue sections.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: M13
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: -20°C

MAP2 protein structure - 13-1500 shown in red.

Supportive validation

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Experimental details: Immunofluorescence analysis of MAP2 was done on 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MAP2 (M13) Mouse Monoclonal Antibody (Product # 13-1500) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

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Experimental details: Immunofluorescent analysis of PSD-95 (green) and MAP2 (red) on rat primary cortical neurons cultured for 28 days in the B-27 Plus Neuronal Culture System (Product # A3653401). At day 28 the cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.1% triton x-100 for 30min, and blocked with 1% BSA for 30 min at room temperature. Cells were stained with anti-PSD95 antibody (Product # 51-6900) at a dilution of 1:200, and anti-MAP2 (Product # 13-1500) at a dilution of 1:400, in 1% BSA staining buffer, overnight at 4C, and then incubated with Alexa Fluor 488 conjugated donkey anti-rabbit (Product # A-21206) and Alexa Fluor 594 donkey anti-mouse (Product # A-21203) antibodies at a dilution of 1:1000 for 30 min. at room temp. Wash 3 times with DPBS. Stain with DAPI for nucleus.

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Experimental details: NULL

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Experimental details: NULL

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Experimental details: Figure 1 Confocal microscopic images of immunohistochemical staining of differentiated SH-SY5Y neural cells following differentiation in staurosporine . Abbreviations: TUJ1 (Neuronal Class III beta-Tubulin); MAP-2 (microtubule associated protein 2); NF68 (neurofilament 68); alpha SYN (alpha-synuclein); VMAT (vesicular monoamine transporter) TH (tyrosine hydroxylase); DAT (dopamine transporter); D2 (D2 dopamine receptor); Synap (synaptophysin).

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Experimental details: Figure 1 Marker expression of NTERA2 (NT2) cells and ATRA differentiated NTERA2 neurons (NT2N). NT2 cells (A-C) co-express GFAP and SSEA3 with GFAP staining confined to the nucleus (A). NT2 cells also co-express GFAP and beta-tubulin III (BT3) which are both mainly confined to the nucleus (B). Nestin with GFAP are co-localized in NT2 cells (C). NT2N assembled themselves into cell clusters which co-expressed GFAP and BT3 found within the cytoplasm and neurites (D). Arrow heads point to cells that appear as radial glia cells which only display GFAP staining (D). NT2N also express neural markers TAU and MAP2 with MAP2 confined to dendrites (tapering morphology shown by arrow heads) and TAU not displaying restriction to axons. In merge panel insert shows close-up view of neurite (E). Synapsin 1 staining is seen as punctate along neurite projections co-expressing BT3 (F). Photomicrographs were obtained at 100X (A-C; scale bar 200 um) and 200X (D-F; scale bar 100 um).

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Experimental details: Fluorescently stained images of neurons, which have been differentiated from hNSCs after a-d) 25 DIV and e) 10 Post Conception Weeks (PCWs). (a) Spontaneous differentiation on H-NDs, b) spontaneous on O-NDs, c) Spontaneous Differentiation (SD) on the glass Control. d,e) Induced Differentiation (ID) into neurons on TCPS control after 25 DIV and 10 PCWs respectively. Neurons have been stained using MAP2 (green), NF200 (red), and Hoechst (blue). All scale bars 100 um, magnification 40x.

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Experimental details: FIGURE 5 Distribution of the MAP2 (green signals) in DPSC-derived neuronal-like stem cells grew for 7 days on glass (A) and ECM (B) support. MAP2 was detected by a mouse primary antibody followed by a FITC-conjugated goat anti-mouse secondary antibody. Nuclei were stained with DAPI (C-D) and a merge was made (E-F) . Images were captured using lasers with wavelenght of 405 and 488 nm. Scale bar corresponds to 20 mum.

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Experimental details: FIGURE 9 Western blot assay. The data are expressed as mean +- standard deviation (*** P < 0.001 vs. Control as determined by Student''s t -test) from at least three independent experiments.

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Experimental details: FIGURE 1 Morphology of iPSC/NSC-derived neurons. Neurite networks of healthy donor control (Ct1 and Ct2) and patient (Pt1 and Pt2) neurons were revealed by phase contrast imaging (A) and immunofluorescence imaging of cells stained with antibodies to cytoskeletal proteins beta-3-Tubulin (B) and MAP2 (C) . The length (D) , number (E) and thickness (F) of neurites were measured from phase contrast images and quantified. There were no significant differences in either the number or the length of neurites between control or patient-cells. Specifically, the average neurite length (D) of different cells was: Ct1, 93.2 +- 8.4 mum; Ct2, 94.6 +- 11.6 mum; Pt1, 96.1 +- 10.1 mum; and Pt2, 86.2 +- 7.0 mum ( p = 0.882; n = 5). The average number (E) of neurites per image was: Ct1, 225.8 +- 46.4; Ct 2, 247.2 +- 54.6: Pt1, 236.0 +- 42.4; and Pt2, 225.4 +- 39.8 ( p = 0.985; n = 5). The average neurite thickness (F) in Ct1 was 5.4 +- 0.3 mum, Ct2 was 5.8 +- 0.2 mum, Pt1 was 4.5 +- 0.2 mum, and Pt2 was 4.3 +- 0.2 mum ( p = 0.03 for Ct1 vs Pt1; p = 0.00045 for Ct2 vs Pt1; p = 0.005 for Ct1 vs Pt2; p = 0.00004 for Ct2 vs Pt2; n = 10 primary neurites measured in 5 phase contrast images for all conditions, totaling 50 neurites measured per condition). * Indicates a significance level of p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. (G) Number of DAPI-Positive Cells Bar Graph. DAPI staining of nuclei revealed fewer patient neurons adhered to the culture dish. Number of DAPI-positive cel

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Experimental details: FIGURE 5 (A) Calcium imaging data. Top panels show raw traces of spontaneous calcium transients recorded from neurons of control donors and patients. Bottom bar graphs are quantitative data of the mean peak amplitudes and spiking frequency of calcium transients. Our data show that neurons from patients (Pt1 and Pt2) exhibited significant changes in either the amplitude or the frequency of calcium transients with the inability to return to basal levels compared to their matched donor controls (Ct1 and Ct2). Arrows indicate basal levels between calcium transcients. (B) Adhesion activity of neurons. Control (Ct1 and Ct2) and patient-derived (Pt1 and Pt2) neurons (6-days after differentiation from NSC) were plated onto Geltrex coated 8-well glass slide Chamber and incubated at 37degC for 30 min, the dishes were flipped to remove non-adherent cells. The adherent neurons were stained with an antibody specific to MAP2 and DAPI and MAP2-expressing cells were counted. Percentage of adherent (flipped) cells was calculated based on un-flipped cells. Patient-derived cells exhibited statistically less adhesion than control cells Ct1 and Ct2: Pt1 ( p < 0.05) and Pt2 (

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Experimental details: Figure 1 Kinin B1 receptor (B1R), A Disintegrin And Metalloprotease 17 (ADAM17), and angiotensin converting enzyme 2 (ACE2) expression in primary hypothalamic neurons. Representative photomicrographs showing immunofluorescence staining for neuron-specific marker microtubule associated protein 2 (MAP-2) (Green) in primary neurons from wild-type ( A ) and B1R knockout mice ( B ) cultured for 10 days. Primary neurons from wild-type mice pups showed immunopositivity for the presence of B1R ( C ), while B1R immunoreactivity was absent in neurons with B1R deletion ( D ). Representative triple immunostaining revealed that kinin B1R ( E ) and ADAM17 ( F ) are expressed in primary hypothalamic neurons expressing nuclear DAPI staining in blue ( G ), and merged image ( H ) shows the colocalization of B1R and ADAM17. Representative triple immunostaining showing ACE2 and ADAM17 expression and co-localization in wild-type neurons ( I - L ) and in B1R knockout neurons ( M - P ).

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Experimental details: Fig 2 CUL9 KO clones differentiate to hNPCs and early born cortical neurons. ( A ) Protocol for neuronal differentiation of hPSCs. Diagram above in vitro differentiation roughly correlates to in vivo corticogenesis with differentiation protocol. Exact cell population at stages of differentiation often varies between differentiations. NPC = neuroepithelial progenitor cells; PP = preplate; vRGC = ventricular radial glia cells; IPC = intermediate progenitor cells; SP = subplate; CP = cortical plate; oRGC = outer radial glia cells; Ast = astrocyte ( B ) CUL9 protein levels increase over the course of cortical neuron differentiation. Western blot analysis of lysates obtained from WT neurons differentiated for ten days and 135 days show gradual increase in CUL9 protein levels compared to WT hPSCs. APC7 levels increase during initial neuronal induction, and then remain relatively stable throughout maturation. n = 3. ( C ) CUL9 mRNA levels increase over the course of cortical differentiation. RNA isolated from iPSCs, D7 cortical differentiation, and D25 cortical differentiation were analyzed by RT-qPCR. Error bars +/- SD. iPSC and D7 n = 2. D25 n = 3. Key hNPC ( D ) and neuronal markers ( E ) are appropriately localized and expressed in CUL9 KO cells. hNSCs were fixed with 4% PFA and stained with key transcription factor PAX6 (red, Alexa 546) and NESTIN (purple, Alexa 647), and Hoechst, marking nuclei (blue). hNPCs were fixed and stained with neuronal markers MAP2 (purple, Alexa 647)

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Experimental details: Figure 5 The effect of extracellular choline uptake inhibition on neurite outgrowth in hNSCs. ( A ) Cultivation in the control differentiation medium (0 muM HC-3) for 7 days. Staining shows microtubule-associated protein 2 (MAP2) (red) and DAPI (blue). Scale bar: 200 mum. ( B ) Cultivation in the differentiation medium with 250 muM HC-3 for 7 days. Staining shows MAP2 (red) and DAPI (blue). Scale bar: 200 mum. ( C ) Comparison of the neurite length in various HC-3 concentrations medium. The neurite length was measured in randomly chosen cells (30 cells) by tracing individual neurites. **** p < 0.0001 denotes statistical significance vs. 0 muM HC-3 group using Dunnett multiple comparison test. ns means not significant.

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Experimental details: 1 FIGURE alpha-Synuclein preformed fibrils (PFF) preparation, Characterization, and SH-SY5Y neuroblastoma cell differentiation. (A) Characterization of purified alpha-syn fractions on 15% SDS-PAGE eluted with different NaCl concentrations. (B) Western blot of purified alpha-syn. Western blot analysis of purified alpha-syn, primary antibody: H3C And secondary: Anti-mouse HRP conjugated antibody. (C) Thioflavin-T assay of alpha-synuclein (Red Dots) aggregation. The data represent +-SEM of three independent replicates. (D) Representative fluorescence microscopy image of alpha-synuclein fibrils. Scale bar indicates 5 mum: (E) Representative AFM image of alpha-synuclein fibrils. The scale bar indicates 500 nm. (F-K) show the confocal microscopy images represented as maximum z projection (F-G), Phalloidin (F-Actin) staining in undifferentiated SH-SY5Y cells at day 0, and differentiated neurons on day 21, respectively. (I and J) FM 1-43 dye staining in undifferentiated SH-SY5Y cells at day 0 and differentiated neurons on day 21, respectively. The cells were stained with FM 1-43FX (5 mug/ml) dye for 15 min at 37degC, followed by cell fixation with 4% PFA. (H and K) shows the immunofluorescence analysis done on differentiated neurons on day 21 for Tau, Synaptophysin (SP11) and Microtubule associated protein (MAP2). Mature neurons confirmation was done against Tau, SP11 and MAP2 at 1:250 dilution for 3 h followed by secondary antibody labeling with Goat anti-Rabbit IgG (H + L) Highly C

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Experimental details: FIGURE 3 LC3B lipidation and punctate formation in cortical neurons due to high glucose and PGRN. (A) Western blot analysis of LC3 lipidation (i.e., LC3-II:I ratio) in primary cortical neurons after 72 h of treatment. The LC3-II:I ratio decreased from control to high glucose (1.000 +- 0.208 AU to 0.464 +- 0.092 AU), an effect that appeared unaffected when treated with PGRN alongside high glucose (0.474 +- 0.076 AU). No difference was observed between control and PGRN-treated samples (0.938 +- 0.309 AU). N = 9 samples. (B) Immunofluorescence analysis of LC3B puncta expression in neurons after 72 h of treatment. The amount of punctate expression trended toward an increase due to PGRN (from 1.000 +- 0.069 AU, to 1.183 +- 0.077 AU). This difference was significant under high-glucose conditions (from 0.916 +- 0.045 AU, to 1.190 +- 0.097 AU). N = 11-29 cells. (C) Representative immunofluorescence images of primary neurons after 72 h of treatment, with blue as DAPI, green as MAP2, and red as LC3B. Scale bar, 10 mum. (D) Immunofluorescence of LC3B puncta expression in astrocytes after 72 h of treatment increased significantly due to high glucose as well as PGRN treatment (from 1.000 +- 0.0374 AU to 1.512 +- 0.087 AU and 2.792 +- 0.099 AU, respectively). This decreased to control levels under HG + PGRN treatment, to 0.899 +- 0.049 AU. N = 12-29 cells. (E) Representative immunofluorescence images of primary astrocytes after 72 h of treatment, with blue as DAPI, green as GFAP, and red a

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Experimental details: FIGURE 4 LAMP2A protein levels and punctate localization in cortical neurons and astrocytes due to high glucose and PGRN treatment. (A) Total protein LAMP2A levels are unchanged among treatment groups in primary cortical neurons treated for 24 h. N = 6 samples. (B) Perinuclear LAMP2A punctate formation increased in neurons under HG (from 1.000 +- 0.137 AU to 1.463 +- 0.108 AU), an effect that was attenuated with PGRN (from 1.031 +- 0.083 AU to 1.246 +- 0.096 AU). N = 12-27 cells. (C) Representative immunofluorescence images of primary neurons after 72 h of treatment, with blue as DAPI, green as MAP2, and red as LAMP2A. Scale bar, 10 mum. (D) Perinuclear LAMP2A punctate formation appeared to show a trend toward an increase due to PGRN but was not statistically significant ( F = 1.825, p = 0.151). N = 16-22 cells. (E) Representative immunofluorescence images of primary astrocytes after 72 h of treatment, with blue as DAPI, green as GFAP, and red as LAMP2A. Scale bar, 10 mum. ** p < 0.01.

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Experimental details: Fig. 1. Gene expression changes in cultured cortical neurons after bisphenol-A (BPA) exposure. (A,B) Cell viability: WST-8 assay. Histograms represent the percentage, with respect to control cells, of viable cells after exposure to 0.01, 0.1, 1, 10, 100 or 1000 muM BPA for 24 h or 48 h. Statistical significance was determined by two-way ANOVA with Bonferroni correction. Data are shown as relative changes versus controls. NS, not significant; * P

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Experimental details: Fig. 2. Abnormal neurite architecture in cultured cortical neurons after BPA exposure. (A) BPA initially suppressed neurite outgrowth in cultured cortical neurons. At 5 days in vitro (DIV5), cultured cortical neurons were treated with saline or BPA (100 muM) for 24 h. Neurites were assessed by immunostaining using an anti-MAP2 antibody. Axons were assessed by immunostaining using an anti-TUJ1 antibody. Scale bar: 10 mum. (B) Quantification of the number of primary neurites in each condition. n =60 neurons for control; n =60 neurons for the BPA-exposed condition. (C) Quantification of the length of primary neurites in each condition. n =60 neurons for control; n =60 neurons for the BPA-exposed condition. (D) Quantification of the length of axons in each condition. n =60 neurons for control; n =60 neurons for the BPA condition. Statistical significance was determined by one-way ANOVA with Bonferroni correction. Data are shown as relative changes versus controls. ** P

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Experimental details: Figure 7. Studies of GPER expression in hippocampal and cortical cultures and tissues. GPER expression and localization in cultured hippocampal and cortical neurons 72 HIC were measured using immunofluorescent and confocal microscopy techniques. A , Representative fluorescence images showing the localization of GPER (red) in cell bodies (arrowheads) and neurites (arrows). Cell nuclei are labeled with DAPI (blue) and neurites are labeled with MAP2 (green). B , RT-qPCR measurement of the relative GPER mRNA level in samples derived from hippocampal and cortical culture at 72 HIC shows a significantly (unpaired Student's t test, p < 0.01, n = 5 replicates) higher GPER mRNA level in cortical than hippocampal cultures. C , Western blot measurements of GPER protein expression in ex vivo hippocampal and cortical tissues from individual E18 rat brains ( n = 10) reveals two protein species with mass sizes of ~50 and 42 kDa ( i ). Statistical analysis shows that expression of GPER ~50 kDa is significantly higher in cortical than in hippocampal tissues ( ii ), while GPER ~42 kDa is slightly, but not significantly, higher in cortical than in hippocampal tissues ( iii ). Unpaired Student's t test; ** p < 0.01. The specificity of the GPER antibody used for this study was validated by immunocytochemistry and Western blotting. The validation results are shown in Extended Data Figure 7-1 .

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Experimental details: Fig. 2 dCas9-Dnmt3a-mediated APP methylation enhanced neuroprotection in APP -KI mouse primary neurons. a Quantitative real-time PCR analysis of APP expression in WT and APP -KI mouse primary neurons treated with - 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean +- SEM ( n = 3). ** P < 0.01, two-sided Student's t -test. b , c Western blot analysis of APP in WT and APP -KI mouse primary neurons treated with - 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean +- SEM ( n = 3). ** P < 0.01, two-sided Student's t -test. d Bisulfite sequencing analysis of the promoter region of APP in APP -KI mouse primary neurons treated with - 189 sgRNA and dCas9-Dnmt3a. e Quantification of methylated amplicons. In 3 independent experiments, 10 to 30 sequencing data were measured in total. Data are expressed as mean +- SEM. ** P < 0.01, two-sided Student's t -test. f Immunostaining of Tuj1 (green), NeuN (red), and DAPI (blue) in APP -KI mouse primary neurons treated with - 189 sgRNA and dCas9-Dnmt3a. g The number of Tuj1-positive neurons in f . Data are expressed as mean +- SEM ( n = 3). ** P < 0.01, two-sided Student's t -test. h Quantification of neurite length. In 3 independent experiments, over 100 neurites were measured. Data are expressed as mean +- SEM ( n = 3). ** P < 0.01, two-sided Student's t -test. i Map2 (green), Abeta42 (red), and DAPI (blue) immunostaining in APP -KI mouse primary neurons treated with - 189 sgRNA and dCas9-Dnmt3a. j Percentage of Abeta42+/Map2+ cells fr

Supportive validation

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Experimental details: Figure 3. Fluorescence-activated cell sorting analysis of MAP-2, vWF, ALP and PPARgamma-2 following culture of rat mesenchymal stem cells for 10 days in differentiation medium subsequent to exposure to NG and 72 h and 10 days SMG. The gray area is the isotype control. The results represent three independent experiments. MAP-2, microtubule-associated protein 2; vWF, von Willebrand factor; ALP, alkaline phosphatase; PPARgamma2, peroxisome proliferator-activated receptor gamma2; NG, normal gravity; SMG, simulated microgravity.

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Experimental details: Figure 5 Photomicrographs of P1 laminar progenitor cells cultured in adipogenic (A) , osteogenic (B) , neurogenic (C) or stromal (D-F) medium and stained with oil red O (A,D) , alizarin red (B,E) or anti-map2, anti-mouse IgG-488 (green, C,F ). DAPI nuclear stain (blue, C,F ). Magnification = 20X, Bar = 100 mum.

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Experimental details: Flow cytometry histogram showing MAP2 labeling of P1 equine hoof progenitor cells from an unaffected hoof cultured in stromal (Stromal MAP2) or neurogenic medium (Neurogenic MAP2), and autofluorescence of unlabelled cells cultured in neurogenic medium (Neurogenic Auto, A ). Percentage (mean +- SEM) of P1 hoof progenitor cells from unaffected hooves expressing MAP2 after culture in stromal (Stromal) or neurogenic (Neurogenic) induction medium (B) .