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遺伝子ノックアウト細胞株 検証済

Anti-Ezrin 抗体 [3C12] (ab4069)

Reviews (8)Q&A (5)References (62)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)
  • Immunohistochemistry (Frozen sections) - Anti-Ezrin antibody [3C12] (ab4069)
  • Western blot - Anti-Ezrin antibody [3C12] (ab4069)
  • Western blot - Anti-Ezrin antibody [3C12] (ab4069)
  • Flow Cytometry (Intracellular) - Anti-Ezrin antibody [3C12] (ab4069)

Key features and details

  • Mouse monoclonal [3C12] to Ezrin
  • Suitable for: WB, IHC-Fr, Flow Cyt (Intra), IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Ezrin antibody [3C12]
    Ezrin 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [3C12] to Ezrin

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, IHC-Fr, Flow Cyt (Intra), IHC-Pmore details

  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: Rat, Hamster, Cow, Pig, Kangaroo, Monkey

  • 免疫原

    Recombinant fragment corresponding to Human Ezrin aa 350-600.

  • ポジティブ･コントロール

    • Lung.

  • 特記事項

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab4069の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (5)
1/100 - 1/750. Predicted molecular weight: 69 kDa.
IHC-Fr (1)
1/50 - 1/100.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 0.025 - 1 µg/ml.
特記事項
WB
1/100 - 1/750. Predicted molecular weight: 69 kDa.
IHC-Fr
1/50 - 1/100.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use a concentration of 0.025 - 1 µg/ml.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    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.

  • 配列類似性

    Contains 1 FERM domain.

  • 発生段階

    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.

  • 翻訳後修飾

    Phosphorylated by tyrosine-protein kinases.

  • 細胞内局在

    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

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)

    IHC image of Ezrin staining in a section of formalin-fixed paraffin-embedded normal human lung performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab4069, 0.025ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [3C12] (ab4069)

    IHC image of Ezrin staining in mouse lung formalin fixed paraffin embedded tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab4069, 1µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab97040, 1/1000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

  • Immunohistochemistry (Frozen sections) - Anti-Ezrin antibody [3C12] (ab4069)
    Immunohistochemistry (Frozen sections) - Anti-Ezrin antibody [3C12] (ab4069)

    IHC image of ab4069 staining in 10% formaldehyde fixed frozen tissue section of human placenta.

    Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with ab4069 (1μg/ml dilution) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647)) and DAPI for 1 hour at room temperature.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab4069. 

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

  • Western blot - Anti-Ezrin antibody [3C12] (ab4069)
    Western blot - Anti-Ezrin antibody [3C12] (ab4069)
    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.

  • Western blot - Anti-Ezrin antibody [3C12] (ab4069)
    Western blot - Anti-Ezrin antibody [3C12] (ab4069)This image is courtesy of an anonymous Abreview
    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

  • Flow Cytometry (Intracellular) - Anti-Ezrin antibody [3C12] (ab4069)
    Flow Cytometry (Intracellular) - Anti-Ezrin antibody [3C12] (ab4069)

    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.

参考文献 (62)

ab4069 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab4069 は 62 報の論文で使用されています。

  • Crespo-García T  et al. A Cantú syndrome mutation produces dual effects on KATP channels by disrupting ankyrin B regulation. J Gen Physiol 155:N/A (2023). PubMed: 36287534
  • Marcadis AR  et al. Rapid cancer cell perineural invasion utilizes amoeboid migration. Proc Natl Acad Sci U S A 120:e2210735120 (2023). PubMed: 37075074
  • Ma L  et al. Ezrin promotes esophageal squamous cell carcinoma progression via the Hippo signaling pathway. Open Life Sci 18:20220743 (2023). PubMed: 37791063
  • Hausrat TJ  et al. Monoallelic loss of the F-actin-binding protein radixin facilitates startle reactivity and pre-pulse inhibition in mice. Front Cell Dev Biol 10:987691 (2022). PubMed: 36518539
  • Yao Q  et al. Long non-coding RNA taurine up regulated 1 promotes osteosarcoma cell proliferation and invasion through upregulating Ezrin expression as a competing endogenous RNA of micro RNA-377-3p. Bioengineered 13:1767-1778 (2022). PubMed: 35012433
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-10 of 13 Abreviews or Q&A

Western blot abreview for Anti-Ezrin antibody [3C12]

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Muscle)
Gel Running Conditions
Reduced Denaturing (GEL 4-12%)
Loading amount
40 µg
Specification
Muscle
Blocking step
intercept blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Oct 24 2023

Western blot abreview for Anti-Ezrin antibody [3C12]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - other (T1 L1 L2)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
40 µg
Specification
T1 L1 L2
Blocking step
intercept blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Oct 24 2023

Immunohistochemistry (Frozen sections) abreview for Anti-Ezrin antibody [3C12]

 Below Average
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (T1 L1 L2)
Permeabilization
Yes - triton 0.5%
Specification
T1 L1 L2
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MRS. Nathalie Da Silva

Verified customer

投稿 Oct 24 2023

Immunocytochemistry/ Immunofluorescence abreview for Anti-Ezrin antibody [3C12]

 Good
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Sep 11 2014

Western blot abreview for Anti-Ezrin antibody [3C12]

 Average
Abreviews
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Aug 01 2014

Western blot abreview for Anti-Ezrin [3C12] antibody

 Good
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jul 17 2014

Western blot abreview for Anti-Ezrin [3C12] antibody

 Good
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jul 16 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Ezrin antibody [3C12]

 Average
Abreviews
abreview image
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%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Dec 07 2006

Question

Dear Sir/Madam,

I am currently planning to carry out an immunohistochemical pilot study using ezrin antibody in oral squamous cell carcinoma, including a sample of about 15 slides. Therefore I kindly request you to provide me the following details regarding the antibody:

1) Is ezrin antibody expression with immunohistochemistry been proven by your company?
2) Is the ezrin antibodyavailablein prediluted ready-to-use form for immunohistochemistry?
3) If prediluted form is available, what quantity of the product is available from your company?
4) If only concentrated form of antibody is available, what is the dilution factor for this product when used for IHC - paraffin?
5) your website (abcam) shows the product Anti-Ezrin antibody [3C12] (ab4069) is available in 500 microlitres. Is it a prediluted form? If so, what are the other quantities of the product available?
6) If the given quantity of ezrin antibody is diluted with the specified dilution factor, how much final quantity of the product will i be able to get?
7) What is the cost of the antibody?

Hope you provide me the details required and do the needful so that I can purchase the antibody as soon as possible.
Awaiting your reply..

Thanking you Sir/Madam,

Yours sincerely,

Read More
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/Ezrin-antibody-SPM244-prediluted-ab18057.html (or use the following: https://www.abcam.com/Ezrin-antibody-SPM244-prediluted-ab18057.html).

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

Question

Dear Sir/Madam, For my project I wish to study the expression of AKAP proteins, including Ezrin. However, I have learned that antibodies for these proteins are difficult to use (without overexpression). Therefore, I wish to try new antibodies, I was hoping that you could have some kind of introductory discount to try thisAnti-Ezrin antibody [3C12] (ab4069)? I would love to hear from you. Thank you in advance. Kind regards,

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

1-10 of 13 Abreviews or Q&A

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