MA5-12826

antibody from Invitrogen Antibodies

Targeting: MAP2 MAP2A, MAP2B, MAP2C

Provider product page for MA5-12826

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [35]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [4]
Immunohistochemistry [2]
Other assay [10]

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Product number: MA5-12826 - Provider product page
Provider: Invitrogen Antibodies
Product name: MAP2 Monoclonal Antibody (AP18)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-12826 has been successfully used in immunofluorescence analysis of Map2 in human glutamate neurons derived from iPSCs.
Antibody clone number: AP18
Concentration: 0.2 mg/mL

MAP2 protein structure - MA5-12826 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of microtubule-associated protein 2 (MAP2, green) in cultured primary cortical neurons at different developmental stages (day 1 and day 14). Primary cortical neurons were isolated and cultured using the Primary Neuron Isolation Kit (Product # 88280). Neurons were fixed with 4% paraformaldehyde, permeablilized with 0.1% Triton X-100 in HBSS for 10 minutes at room temperature, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were probed with a MAP2 monoclonal antibody, clone AP18 (Product # MA5-12826) at a dilution of 1:500 for 2 hours at room temperature (or overnight at 4°C), washed with HBSS, and incubated with a DyLight 488 goat anti-mouse IgG (Product # 35502) secondary antibody at a dilution of 1:500 for 1 hour at room temperature (panels A and B). In panel C, cells were also probed with a glial fibrillary acidic protein (GFAP) polyclonal antibody at a dilution of 1:500 for 2 hours at room temperature (or overnight at 4°C), washed with HBSS, and incubated with a DyLight 680 goat anti-rabbit IgG (Product # 35568) secondary antibody at a dilution of 1:500 for 1 hour at room temperature. Images were taken at 20X magnification on a Carl Zeiss microscope (AxioVision Rel. 4.7).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of MAP2 a, b, c (green) in SH-SY5Y cells either left untreated (left panel) or treated with 10uM all-trans retinoic acid for 72 hours (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a MAP2 a, b, c monoclonal antibody (Product # MA5-12826) at a dilution of 1:200 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or a ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of MAP2 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MAP2 Monoclonal Antibody (AP18) (Product # MA5-12826) at 1:500 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents Hep G2 cells having no expression of MAP2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Map2 (green) in human glutamate neurons derived from iPSCs. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.2% Triton X-100 in PBS for 10 minutes, and blocked with 5% donkey serum in PBS for 15 minutes at room temperature. Cells were stained with a Map2 monoclonal antibody (Product # MA5-12826) diluted at 1:1000 in 5% donkey serum overnight at 4°C, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for 1 hour at room temperature (green). Note: Data courtesy of Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Effect of naringenin (Nar) on senescence of mouse neural stem cells (mNSCs). ( A ) p16 ink4a mRNA expression was measured by qRT-PCR in 23M-NSCs with and without treatment of 6.8 mug/mL Nar for 48 h. ( B ) Cell cycle phase distributions of 23M-NSCs treated with Nar for 48 h and control cells. ( C ) The relative telomere length of 23M-NSCs increased significantly with Nar treatment. ( D ) Immunofluorescence Ki67 staining of mNSCs treated with Nar for 48 h, with DAPI nuclear labeling. ( E ) Quantification of ( D ). ( F ) Representative fields of MAP2 and GFAP immunofluorescence staining of cultured mNSCs after control and Nar treatment. ( G ) Quantification of ( F ). Data are presented as the mean +- SD of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.0001 compared with untreated cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Characterization of idNs. ( A ) Representation of Na V 1.1 channel. The star in segment 1 of domain I shows the localization of the mutated aminoacid (Met145Thr). The relative missense mutation c.434T > C is found in the exon 3 of the translated sequence. ( B ) Bright-field images of idNs from WT and SCN1A M145T -iPSCs (20x magnification). ( C ) Differentiated idNs show high expression levels of neuronal specific genes such as MAP2 , NEFM , NEFL, SYP and PSD95 compared to their undifferentiated counterparts (iPSCs). GAPDH was used as a housekeeping control. Data are presented as mean +- SEM of three biological replicates (black dots), * p < 0.05, ** p < 0.01, *** p < 0.001, t -test has been calculated vs. expression in iPSCs. ( D ) Immunostaining of neuronal markers TUBB3 (neurites marker), MAP2 (cell body and dendrites marker), and NEFH (axonal marker) in WT and SCN1A M145T idNs. DAPI nuclear counterstain is shown in all images in blue (63x magnification).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Types of interneurons generated from iPSCs. Immunofluorescence analysis of idNs stained with antibodies against interneuronal subtypes markers ( A ) somatostatin (SST), ( B ) calretinin (CALB2), and ( C ) calbindin (CALB1). In each group of images, WT cells are shown in the upper panel, while SCN1A M145T idNs are shown in the lower panel. White arrows in the merged images indicate neurons expressing the interneuronal makers indicated (63x magnification). ( D - F ) Quantification of percentage of MAP2 + neurons co-expressing interneuronal markers immunostained in panels ( A - C ). About 9-1% of idNs express SST and CALB2, while CALB1 is present in less than 1% percent of neurons. At least 200 cells were counted for each bar, and data are presented as mean +- SEM of two independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Reconstruction of the neurovascular unit in the Brain-Chip (A) Schematic illustration of the Brain-Chip, a two-channel microengineered chip including iPSC-derived brain endothelial-like cells cultured on all surfaces of the bottom channel, and iPSC-derived Glutamatergic and GABAergic neurons, primary human brain astrocytes, pericytes, and microglia on the surface of the top channel. (B) Confocal images of the cell coverage in the brain channel on day 7 of culture. Top image: Immunofluorescence staining of the brain channel including MAP2 (green), GFAP (magenta), NG2 (red), and DAPI (blue). Bottom images: Representative merged confocal image of the brain channel on culture day 7, stained for neurons (MAP2, green), astrocytes (GFAP, magenta, IBA1, yellow), and pericytes (alphaSMA, red) (bar, 50 mum). (C) Representative immunofluorescent staining for s100beta (red) and GLAST (green) (bar,100 mum). (D) FACS analysis of cell-specific markers of microglia: Total population of microglia within the brain channel (gray), CD11b-positive population (magenta), CD45-positive population (magenta), quantification of CD11b:CD45-positive cells. (E) Representative merged confocal image of the brain channel co-stained with VGAT (green) for GABAergic neurons and VGlut1 (red) for Glutamatergic neurons (bar, 100 mum). (F) Immunofluorescence staining of the brain channel including MAP2 (green) and SYP (red) (bar, 100 mum). (G) Levels of secreted glutamate in the brain channel on culture days 5, 6,

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4--figure supplement 1. The number of axons is not affected by Drp1 ABCD knockdown. Cultured hippocampal neurons were transfected at 3 weeks with the indicated shRNA plasmids carrying GFP as a cytosolic marker. Cells were subjected to immunofluorescence microscopy with anti-MAP2 antibodies. Arrowheads indicate axons, and arrows indicate dendrites. Bar, 20 um.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Generation and validation of viral vectors for Nrg1 expression. (a) Schematic representation of the constructs for the expression of Nrg1-ICD (including the nuclear localization signal (NLS)) and Nrg1-FL both fused to GFP. The schema of the vector for the production of AAV particles to express Nrg1 under human Synapsin promoter is depicted on the right. CRD: cysteine-rich domain; EGF: epithelial growth factor domain; TM: transmembrane domain; ITR: inverted terminal repeat; ORF: open reading frame; hGH PA: human growth hormone polyadenylation signal. (b-j) Immunofluorescent labeling of primary cortical neurons infected to express GFP, Nrg1-ICD, or Nrg1-FL and fixed at D14. The neurons were labeled with DAPI and microtubule-associated protein 2 (MAP2) to visualize neuronal dendrites. The boxed area in (b, e, h) depicts the area magnified in (d, g, j). The dotted lines delimit the nuclei to highlight the specific localization of Nrg1 constructs. Scale bar, 50 mu m.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Western blot results of neuroblastoma cell protein expression under control, OGD/R conditions, and OGD/R conditions treated with exogenous MIF or MIF antagonist ISO-1. (A) The expression levels of immature and mature BDNF and MAP2 in the MIF group were significantly higher than those in the OGD/R group (immature BDNF: P < 0.05; mature BDNF: P < 0.01). (C) The expression levels of Bcl2, Caspase-3, and Bax in the MIF group were significantly lower than those in the OGD/R group (Caspase-3: P < 0.01; Bax: P = 0.02). The expression levels of Bcl2 and MAP2 were significantly higher in the MIF group and lower in the ISO-1 group than those in the OGD/R group (Bcl2: P < 0.01; MAP2: P < 0.05). The expression levels of Caspase-3 and Bax were significantly lower in the MIF group and higher in the ISO-1 group than those in the OGD/R group (Caspase-3: P < 0.01; Bax: P = 0.02). (B and D) The representative bands of each group in western blot are presented. * P < 0.05, ** P < 0.01, vs . OGD/R group; one-way analysis of variance followed by the Bonferroni post-hoc analysis (number of independent cell culture experiments ( n ): immature BDNF, n = 5; mature BDNF, n = 6; Bcl2, n = 6, MAP2, n = 8; Caspase-3, n = 7; Bax, n = 8). BDNF: Brain-derived neurotrophic factor; ISO-1: MIF antagonist; MAP2: microtubule-associated protein 2; MIF: macrophage migration inhibitory factor recombinant; OGD/R: oxygen-glucose deprivation and reperfusion.