Anti-Ezrin (phospho T567) 抗体

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin (phospho T567) antibody (AB47293)

ab47293 staining human normal placenta. Staining is localised to apical membrane and cytoplasmic compartiment.
Left panel : with primary antibody at 1 ug/ml. Right panel : isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may b

• WB

Supplier Data

Western blot - Anti-Ezrin (phospho T567) antibody (AB47293)

The lower band is likely to be Moesin, which has a homologous immunogenic region.

All lanes:

Western blot - Anti-Ezrin (phospho T567) antibody (ab47293)

Lane 1:

Jurkat cell lysate + PMA (125 ng/ml, 30 minutes)

Lane 2:

Jurkat cell lysate

Predicted band size: 69 kDa

Observed band size: 68 kDa,69 kDa

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• WB

Unknown

Western blot - Anti-Ezrin (phospho T567) antibody (AB47293)

All lanes:

Western blot - Anti-Ezrin (phospho T567) antibody (ab47293)

Lane 1:

EGF treated A431 cells, no added peptide

Lane 2:

Untreated A431 cells, no added peptide

Predicted band size: 69 kDa

Observed band size: 69 kDa

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• WB

CiteAb

Western blot - Anti-Ezrin (phospho T567) antibody (AB47293)

Western Blotting using Anti-Ezrin (phospho T567) antibody, ab47293. Publication image from Sato, K. et al., 2015, Nat Commun, 26563429. Legend direct from paper.

Essential roles of TRPV6 in FSS-induced microvilli formation.(a) Apical localization of ezrin in the FSS-exposed BeWo cells. Cells were cultured overnight under static conditions, and then cultured with or without medium perfusion (5 µl min−1) for 1 h. Ezrin (green) localization was observed by immunofluorescence confocal microscopy. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; blue). Serial x–y focal planes (z-sections, 0.88 µm interval in ‘Static', 0.90 µm in ‘Flow') and the stacked images are shown. Scale bar, 20 µm. (b) Time-course phosphorylation of Ezrin (pThr567) and Akt (pSer473) in response to FSS. BeWo cells were cultured overnight under static conditions and exposed to FSS (5 µl min−1) for the indicated times. The cells were lysed, and protein expression level or phosphorylation was analysed by immunoblotting. (c) Inhibition of Akt phosphorylation by BAPTA-AM. Gö6793 (100 nM), BAPTA-AM (10 µM) or buffer control (DMSO) was added to the perfusing medium. (d) Involvement of TRPV6 in the FSS-induced Ezrin phosphorylation. BeWo cells transfected with siRNA were seeded in the chamber area of the device and cultured overnight under static conditions before exposure to FSS for 1 h. β-Actin was used as a loading control. Note that the TRPV6 siRNA oligo (TRPV6 si-#1 or #2), as well as the siRNA pool, shows similar inhibitory effects on the FSS-induced phosphorylation of Ezrin and Akt. (e) Impaired Ezrin localization in the TRPV6 knockdown cells. Ezrin localization in the TRPV6 knockdown cells was analysed by immunofluorescence confocal microscopy. Note that TRPV6 knockdown cells exposed to FSS (5 µl min−1) for 1 h failed to show the microvillous localization pattern of Ezrin as observed in the control siRNA cells under FSS. Scale bar, 20 µm.

false

• WB

CiteAb

Western blot - Anti-Ezrin (phospho T567) antibody (AB47293)

Western Blotting using Anti-Ezrin (phospho T567) antibody, ab47293. Publication image from Sato, K. et al., 2015, Nat Commun, 26563429. Legend direct from paper.

Essential roles of TRPV6 in FSS-induced microvilli formation.(a) Apical localization of ezrin in the FSS-exposed BeWo cells. Cells were cultured overnight under static conditions, and then cultured with or without medium perfusion (5 µl min−1) for 1 h. Ezrin (green) localization was observed by immunofluorescence confocal microscopy. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; blue). Serial x–y focal planes (z-sections, 0.88 µm interval in ‘Static', 0.90 µm in ‘Flow') and the stacked images are shown. Scale bar, 20 µm. (b) Time-course phosphorylation of Ezrin (pThr567) and Akt (pSer473) in response to FSS. BeWo cells were cultured overnight under static conditions and exposed to FSS (5 µl min−1) for the indicated times. The cells were lysed, and protein expression level or phosphorylation was analysed by immunoblotting. (c) Inhibition of Akt phosphorylation by BAPTA-AM. Gö6793 (100 nM), BAPTA-AM (10 µM) or buffer control (DMSO) was added to the perfusing medium. (d) Involvement of TRPV6 in the FSS-induced Ezrin phosphorylation. BeWo cells transfected with siRNA were seeded in the chamber area of the device and cultured overnight under static conditions before exposure to FSS for 1 h. β-Actin was used as a loading control. Note that the TRPV6 siRNA oligo (TRPV6 si-#1 or #2), as well as the siRNA pool, shows similar inhibitory effects on the FSS-induced phosphorylation of Ezrin and Akt. (e) Impaired Ezrin localization in the TRPV6 knockdown cells. Ezrin localization in the TRPV6 knockdown cells was analysed by immunofluorescence confocal microscopy. Note that TRPV6 knockdown cells exposed to FSS (5 µl min−1) for 1 h failed to show the microvillous localization pattern of Ezrin as observed in the control siRNA cells under FSS. Scale bar, 20 µm.

false

• WB

CiteAb

Western blot - Anti-Ezrin (phospho T567) antibody (AB47293)

Western Blotting using Anti-Ezrin (phospho T567) antibody, ab47293. Publication image from Sato, K. et al., 2015, Nat Commun, 26563429. Legend direct from paper.

Essential roles of TRPV6 in FSS-induced microvilli formation.(a) Apical localization of ezrin in the FSS-exposed BeWo cells. Cells were cultured overnight under static conditions, and then cultured with or without medium perfusion (5 µl min−1) for 1 h. Ezrin (green) localization was observed by immunofluorescence confocal microscopy. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; blue). Serial x–y focal planes (z-sections, 0.88 µm interval in ‘Static', 0.90 µm in ‘Flow') and the stacked images are shown. Scale bar, 20 µm. (b) Time-course phosphorylation of Ezrin (pThr567) and Akt (pSer473) in response to FSS. BeWo cells were cultured overnight under static conditions and exposed to FSS (5 µl min−1) for the indicated times. The cells were lysed, and protein expression level or phosphorylation was analysed by immunoblotting. (c) Inhibition of Akt phosphorylation by BAPTA-AM. Gö6793 (100 nM), BAPTA-AM (10 µM) or buffer control (DMSO) was added to the perfusing medium. (d) Involvement of TRPV6 in the FSS-induced Ezrin phosphorylation. BeWo cells transfected with siRNA were seeded in the chamber area of the device and cultured overnight under static conditions before exposure to FSS for 1 h. β-Actin was used as a loading control. Note that the TRPV6 siRNA oligo (TRPV6 si-#1 or #2), as well as the siRNA pool, shows similar inhibitory effects on the FSS-induced phosphorylation of Ezrin and Akt. (e) Impaired Ezrin localization in the TRPV6 knockdown cells. Ezrin localization in the TRPV6 knockdown cells was analysed by immunofluorescence confocal microscopy. Note that TRPV6 knockdown cells exposed to FSS (5 µl min−1) for 1 h failed to show the microvillous localization pattern of Ezrin as observed in the control siRNA cells under FSS. Scale bar, 20 µm.

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