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

Product name

Anti-Ezrin antibody [EPR23353-55]
See all Ezrin primary antibodies
Description

Rabbit monoclonal [EPR23353-55] to Ezrin
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa and A549 cell lysate; Human kidney tissue lysate; Mouse hippocampus and fetal brain tissue; Rat brain lysate; Neuro-2a, C6, PC-12 and NIH/3T3 cell lysate. IHC-P: Mouse kidney and spleen tissue; Rat kidney, liver and spleen tissue. ICC/IF: HeLa and Neuro-2a cells. Flow: HeLa, C6 and Neuro-2a cells. IP: HeLa, C6 and Neuro-2a cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23353-55
Isotype

IgG
Research areas

Alternative Versions
- Anti-Ezrin antibody [EPR23353-55] - BSA and Azide free (ab270525)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab270442 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/500.
ICC/IF		1/50. 100% methanol fixation is recommended.
IP		1/30.
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Predicted molecular weight: 69 kDa.

Notes
Flow Cyt (Intra) 1/500.
ICC/IF 1/50. 100% methanol fixation is recommended.
IP 1/30.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 69 kDa.

Function

Probably involved in connections of major cytoskeletal structures to the plasma membrane. In epithelial cells, required for the formation of microvilli and membrane ruffles on the apical pole. Along with PLEKHG6, required for normal macropinocytosis.
Tissue specificity

Expressed in cerebral cortex, basal ganglia, hippocampus, hypophysis, and optic nerve. Weakly expressed in brain stem and diencephalon. Stronger expression was detected in gray matter of frontal lobe compared to white matter (at protein level). Component of the microvilli of intestinal epithelial cells. Preferentially expressed in astrocytes of hippocampus, frontal cortex, thalamus, parahippocampal cortex, amygdala, insula, and corpus callosum. Not detected in neurons in most tissues studied.
Sequence similarities

Contains 1 FERM domain.
Developmental stage

Very strong staining is detected in the Purkinje cell layer and in part of the molecular layer of the infant brain compared to adult brain.
Post-translational
modifications

Phosphorylated by tyrosine-protein kinases.
Cellular localization

Apical cell membrane. Cell projection. Cell projection > microvillus membrane. Cell projection > ruffle membrane. Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. Localization to the apical membrane of parietal cells depends on the interaction with MPP5. Localizes to cell extensions and peripheral processes of astrocytes (By similarity). Microvillar peripheral membrane protein.
Target information above from: UniProt accession P15311 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 7430 Human
- Entrez Gene: 22350 Mouse
- Entrez Gene: 54319 Rat
- Omim: 123900 Human
- SwissProt: P15311 Human
- SwissProt: P26040 Mouse
- SwissProt: P31977 Rat
- Unigene: 487027 Human
- Unigene: 277812 Mouse
- Unigene: 773 Rat
see all
Alternative names
- Villin 2 ezrin antibody
- CVIL antibody
- CVL antibody
- Cytovillin 2 antibody
- Cytovillin antibody
- DKFZp762H157 antibody
- Epididymis secretory protein Li 105 antibody
- EZR antibody
- EZRI_HUMAN antibody
- Ezrin antibody
- FLJ26216 antibody
- HEL S 105 antibody
- MGC1584 antibody
- p81 antibody
- VIL 2 antibody
- VIL2 antibody
- Villin 2 (ezrin) antibody
- Villin 2 antibody
- Villin-2 antibody
- Villin2 antibody
see all

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.
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).
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.
Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442)

All lanes : Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1 : Mouse hippocampus tissue lysate
Lane 2 : Mouse fetal brain tissue lysate
Lane 3 : Rat brain tissue lysate
Lane 4 : Neuro-2a ( mouse neuroblastoma neuroblast) whole cell lysate
Lane 5 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 7 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 69 kDa

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).
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.
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.
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.
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).
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).
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).
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).
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.
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.
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.
Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442)

All lanes : Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : A549 ( human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : MDA-MB-435S ( human mammary gland ductal carcinoma melanocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 69 kDa

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).
Western blot - Anti-Ezrin antibody [EPR23353-55] (ab270442)

All lanes : Anti-Ezrin antibody [EPR23353-55] (ab270442) at 1/1000 dilution

Lane 1 : Human liver tissue lysate
Lane 2 : Human kidney tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 69 kDa

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

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Developmental stage

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications