Overview

  • Product name
    Anti-Ezrin antibody [3C12]
    See all Ezrin primary antibodies
  • Description
    Mouse monoclonal [3C12] to Ezrin
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-FoFr, WB, ICC/IF, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Cow, Human, Pig, Kangaroo, Monkey
  • Immunogen

    Recombinant fragment, corresponding to amino acids 362-585 of Human Ezrin.

  • Positive control
    • Lung.

Properties

Applications

Our Abpromise guarantee covers the use of ab4069 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
IHC-FoFr Use at an assay dependent concentration. PubMed: 22427845
WB 1/100 - 1/750. Predicted molecular weight: 69 kDa.
ICC/IF Use at an assay dependent concentration. PubMed: 17311105
IHC-P 1/50 - 1/100.
IHC-Fr 1/50 - 1/100.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • 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.
  • Information by UniProt
  • Database links
  • 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

Images

  • All lanes : Anti-Ezrin antibody [3C12] (ab4069) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : EZR (Ezrin) knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : MEF whole cell lysate
    Lane 5 : NIH3T3 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 69 kDa



    Lanes 1 - 5: Merged signal (red and green). Green - ab4069 observed at 81 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab4069 was shown to specifically react with Ezrin in wild-type HAP1 cells as signal was lost in EZR (Ezrin) knockout cells. Wild-type and EZR (Ezrin) knockout samples were subjected to SDS-PAGE. Ab4069 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-Ezrin antibody [3C12] (ab4069) at 1/500 dilution + Mouse bEnd.3 whole cell lysate at 20 µg

    Secondary
    HRP-conjugated goat anti-mouse IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 69 kDa
    Observed band size: 81 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds

    See Abreview

  • Rat lung epithelial microvilli stained with ab4069 (anti-Ezrin) in frozen sections. Review by Marko Nykanen submitted 1 July 2004.
  • ab4069 at 1/100 dilution staining human HeLa cells by ICC/IF. The cells were formaldehyde fixed and blocked with BSA/goat serum prior to incubation with the antibody for 30 minutes. An Alexa-Fluor ® 568 conjugated goat anti-mouse IgG was used as the secondary.

    See Abreview

  • ab4069 staining human lung by IHC-P.
  • Overlay histogram showing SH-SY5Y cells stained with ab4069 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab4069, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human lung adenocarcinoma tissue, staining Ezrin with ab4069.
  • Immunohistochemical analysis of murine retina tissue, staining Ezrin (red) with ab4069.

    Tissue was blocked with blocking solution before incubation with primary antibody (1/50) overnight at 4°C. An AlexaFluor®555-conjugated donkey anti-mouse IgG (1/20) was used as the secondary antibody.

References

This product has been referenced in:
  • Wen Q  et al. Actin nucleator Spire 1 is a regulator of ectoplasmic specialization in the testis. Cell Death Dis 9:208 (2018). WB . Read more (PubMed: 29434191) »
  • Yang YN  et al. miR-204 reverses temozolomide resistance and inhibits cancer initiating cells phenotypes by degrading FAP-a in glioblastoma. Oncol Lett 15:7563-7570 (2018). Read more (PubMed: 29725461) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (PAEC)
Specification
PAEC
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 11 2014

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Non-reduced Denaturing (SDS gel 10%)
Sample
Human Cell lysate - whole cell (Ovarian cancer)
Specification
Ovarian cancer
Treatment
50uM Metformin for 24hr
Blocking step
Milk as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 01 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (HUVEC)
Specification
HUVEC
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 17 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (bEnd.3)
Specification
bEnd.3
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 16 2014

Answer

Thank you for your enquiry. I am pleased to help answer your questions:

1. Any IHC-P testing images we have will be available on the individual online datasheets. Please let me know if there is not one available for the antibody you are interested in, and I can try to obtain this for you.

2. We have one Ezrin antibody tested in IHC-P and human samples which is prediluted:

ab18057 Mouse monoclonal Ezrin antibody [SPM244], prediluted
Tested in: IHC-P
Reacts with: Human, Monkey
https://www.abcam.com/index.html?datasheet=18057 (or use the following: https://www.abcam.com/index.html?datasheet=18057).

3. I am sorry to confirm that ab18057is not in stock at the moment, but stock will be available within to send out 1 - 2 weeksafter your order. however, please note that as you are in India, delivery after this could take more time. I can suggest to contact one of our Indian distributors for more information (please see below).

4. The recommended dilution for IHC-P is usually provided on the individual online datasheet: However, please note that this is a recommendation only and may require some individual optimization.

5. I can confirm that ab4049 is not provided in prediluted form. If an antibody is prediluted, this will be stated.

The online datasheet for ab4049 recommends to dilute at 1:50 - 1:100 for IHC-P.

6. This will depend on which particular antibody you choose, what optimized dilution of antibody you use, and how much you put on each slide.

7. As you are contacting us from India, I can suggest to contact one of our Indian distributors who will be able to provide a price for you. They will also be able to provide an estimate on how long it will take for an item to ship to you.

Central and South India:
Allied Scientific Products
39/19B Gopal Nagar Road
Kolkata-700027
India
Website http://www.alliedscientific.net
Freephone 1800 345 7777
Fax +91 33 24566683 / 24790626
Tel +91 33 24566684/24793109/9039


North India:
NuLife Consultants & Distributors (Pvt.) Ltd.
NuLife Consultants & Distributors (Pvt.) Ltd.
M-52, First Floor,
Lajpat Nagar- Part-II,
New Delhi-110024-India
Tel 011-46112691/ 46112692/ 29836141
Website http://www.nulife.in
Fax 91 - 011-46586216
Email mailto:nulife@bol.net.in
mailto:sales@nulife.in

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your enquiry and happy new year to you. I am sorry to confirm that because we carry over 70,000 products, it is not feasible for us to keep small sample sizes of our products to send to customers. We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase. If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. If you are interested to find out more, please do not hesitate to contact me again by replying to this email prior to purchase for more information. ab4069 Anti-Ezrin antibody [3C12] has been tested and guaranteed in: Applications: WB, ICC/IF, IHC-P, IHC-Fr Species: Mouse, Rat, Hamster, Cow, Human, Kangaroo, Monkey Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Blocking step
BSA with goat serum as blocking agent for 30 minute(s) · Concentration: 10%

Abcam user community

Verified customer

Submitted Dec 07 2006

Question
Answer

Thank you for your enquiry. The concentration is 100 ug/ml. If you have any additional questions, please contact us again.

Read More

Answer

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested in this application. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image). I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

Read More

Question
Answer

The concentration is 100 ug/ml. If you have any additional questions, please contact us again.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up