Anti-Ezrin 抗体 [EPR23353-55]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ezrin with ab270442 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunofluorescent analysis of 100% methanol-fixed HeLa cells labeling Ezrin with ab270442 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

100% methanol fixation is recommended.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Ezrin was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab270442 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270442 at 1/1000 dilution (0.549 μg/ml). VeriBlot for IP Detection Reagent (HRP )(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg.

Lane 2 : ab270442 IP in HeLa whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270442 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 32 secs.

All lanes:

Immunoprecipitation - Anti-Ezrin antibody [EPR23353-55] (ab270442)

Predicted band size: 69 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunofluorescent analysis of 100% methanol-fixed Neuro-2a cells labeling Ezrin with ab270442 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in Neuro-2a cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

100% methanol fixation is recommended.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized C6 (Rat glial tumor glial cell) cells labeling Ezrin with ab270442 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Ezrin with ab270442 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse spleen. The section was incubated with ab270442 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Ezrin with ab270442 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat kidney (PMID : 16522749). The section was incubated with ab270442 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Ezrin with ab270442 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat spleen.The section was incubated with ab270442 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Ezrin with ab270442 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse kidney (PMID : 16522749). The section was incubated with ab270442 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Ezrin with ab270442 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on biliary epithelial cells in rat liver (PMID : 23507543). The section was incubated with ab270442 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling Ezrin with ab270442 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Ezrin was immunoprecipitated from 0.35 mg C6 (rat glial tumor glial cell) whole cell lysate with ab270442 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 270442 1/1000 dilution (0.549 μg/ml). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : C6 whole cell lysate 10 μg.
Lane 2 : ab270442 IP in C6 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270442 in C6 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 84 secs.

The band of 55kDa is endogenously cleaved N-terminus form.

All lanes:

Immunoprecipitation - Anti-Ezrin antibody [EPR23353-55] (ab270442)

Predicted band size: 69 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Ezrin was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab270442 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270442 at 1/1000 dilution (0.549 μg/ml). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a whole cell lysate 10 μg.
Lane 2 : ab270442 IP in Neuro-2a whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270442 in Neuro-2a whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 mins.

All lanes:

Immunoprecipitation - Anti-Ezrin antibody [EPR23353-55] (ab270442)

Predicted band size: 69 kDa

false

• WB

Supplier Data

Western blot - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 6 secs.

The band of 55kDa is the endogenously cleaved N-terminal form.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11124833, 17370041, 10388520).
Negative control : Human liver (PMID : 11124833).

All lanes:

Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 69 kDa

false

• WB

Supplier Data

Western blot - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

Exposure time : 6 secs.

The band of 55kDa is the endogenously cleaved N-terminal form.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11124833, 17370041, 10388520).

Lower expression cell line : MDA-MB-435S (PMID : 17370041).

All lanes:

Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 ( human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MDA-MB-435S ( human mammary gland ductal carcinoma melanocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 69 kDa

false

• WB

Lab

Western blot - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Western blot : Anti-Ezrin antibody [EPR23353-55] ab270442 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 76 kDa in Wild-type A549 cell lysates with no signal observed at this size in EZR knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4 C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human EZR knockout A549 cell line (<a href='/products/cell-lines/human-ezr-knockout-a549-cell-line-ab300934'>ab300934</a>) at 20 µg

Lane 3:

HeLa at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 76 kDa

Observed band size: 76 kDa

false

• WB

Supplier Data

Western blot - Anti-Ezrin antibody [EPR23353-55] (AB270442)

Blocking/diluting buffer and concentration : 5% NFDM/TBST.

Exposure times : Lanes 1-6 : 15 secs; Lane 7 : 3 mins.

The band of 55kDa is the endogenously cleaved N-terminal form.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11124833, 17370041, 10388520).

All lanes:

Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Mouse fetal brain tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Neuro-2a ( mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 7:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 69 kDa

false