Anti-EB3 antibody [EPR11421(B)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EB3 antibody [EPR11421(B)] (AB157217)

Immunohistochemical analysis of paraffin embedded human skeletal muscle tissue labeling EB3 with ab157217 at 1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EB3 antibody [EPR11421(B)] (AB157217)

Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling EB3 with purified ab157217 at a dilution of 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Control : PBS only.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-EB3 antibody [EPR11421(B)] (AB157217)

Intracellular Flow Cytometry analysis of U87-MG (human glioblastoma) cells labeling EB3 with unpurified ab157217 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EB3 antibody [EPR11421(B)] (AB157217)

Immunohistochemical analysis of paraffin embedded human brain tissue labeling EB3 with ab157217 at 1/250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-EB3 antibody [EPR11421(B)] (AB157217)

All lanes:

Western blot - Anti-EB3 antibody [EPR11421(B)] (ab157217) at 1/10000 dilution

Lane 1:

Human cerebellum lysate at 10 µg

Lane 2:

Human fetal brain lysate at 10 µg

Lane 3:

Rat muscle lysate at 10 µg

Predicted band size: 31 kDa

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• WB

CiteAb

Western blot - Anti-EB3 antibody [EPR11421(B)] (AB157217)

EB3 western blot using anti-EB3 antibody [EPR11421(B)] ab157217. Publication image and figure legend from Pchitskaya, E., Rakovskaya, A., et al., 2022, Int J Mol Sci, PubMed 35216391.

ab157217 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab157217 please see the product overview.

EB3 affects PSD-95 and Synapsin protein clustering. (A) The efficiency of EB3 expression level regulation generated in HEK293T cells Lentiviral particles were analyzed by Western blot the primary hippocampal cultures infected with lentiviruses encoding EB3, control RNAi (shControl), or RNAi against EB3 (shEB3) at DIV 7-8 and lysed at DIV 15. Actin was used as a loading control. (B) Quantification of EB3 expression level (normalized to actin levels). The mean density of each band was normalized to an actin signal in the same sample and averaged. n = 4 batches of culture, * : p < 0.05. (C) Area and density of PSD-95 protein clusters for each group of cells shown on panel D. (D) Binarized confocal images of WT hippocampal dendrites and dendrites treated with beta-amyloid, infected with lentiviruses encoding EB3, control RNAi (shControl) or RNAi against EB3 (shEB3) at DIV 7 and fixed at DIV 14 and stained anti-MAP2 (red), anti-PSD95 (blue), and anti-Synapsin (green). Scale bar corresponds to 5μm. (E) Area and density of Synapsin protein clusters for each group of cells shown on panel D. * : p < 0.05, ** : p < 0.01, *** : p < 0.001, **** : p < 0.0001, ns-no statistically significant difference. Tamhane's T3 test for PSD95 clusters area, Kruskal–Wallis test with post-hoc Dunn's test for other data. n ≥ 25 neurons from 3 to 5 batches of cultures.

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