Anti-E2 tag抗体[5E11] (ab977)

Reviews (2)Q&A (3)References (9)

概述

  • 产品名称
    Anti-E2 tag抗体[5E11]
    参阅全部 E2 tag 一抗
  • 描述
    小鼠单克隆抗体[5E11] to E2 tag
  • 宿主
    Mouse
  • 经测试应用
    适用于: IP, WB, ICC/IFmore details
  • 表位
    SSTSSDFRDR (linear)
  • 常规说明
    E2TAG having sequence SSTSSDFRDR or GVSSTSSDFRDR and antibodies specific for the peptide tag are covered by PCT Patent No PCT/EE01/00001 held by Quattromed Ltd. End-user is granted to use the licenced technology solely for internal research purpose only. This antibody should be used in conjunction with the E2 tagging vectors (links to the appropriate datasheet can be found in the Related Products section at the bottom of the datasheet).

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液
    Constituent: 50% Glycerol
  • Concentration information loading...
  • 纯度
    Protein G purified
  • 克隆
    单克隆
  • 克隆编号
    5E11
  • 同种型
    IgG1
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab977 in the following tested applications.

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

应用 Ab评论 说明
IP Use at an assay dependent dilution.
WB 1/1000 - 1/5000. Incubate for 1 hour at room temperature. Background can be reduced by adding NaCl up to a concentration of 1.5M.
ICC/IF 1/1000. Incubate for 1 hour at room temperature.

靶标

  • 相关性
    The E2Tag is a peptide which is derived from the Bovine Pappillomavirus type-1 transactivator protein E2 and consists of 10 amino acids(SSTSSDFRDR). The detection of E2-tagged proteins is based on the binding of mouse monoclonal antibodies specific to the tagged recombinant protein.
  • 别名
    • E2 epitope tag antibody
    • SSTSSDFRDR epitope tag antibody
    • SSTSSDFRDR tag antibody

图片

  • Western blot - Anti-E2 tag antibody [5E11] (ab977)
    Western blot - Anti-E2 tag antibody [5E11] (ab977)
    Cos-7 cell lysates (1µg of total protein per lane) were transfected with pQM vectors encoding two different E2-tagged proteins. Detection is performed with anti-E2Tag antibody (5E11) (1:10 000 in TBST) with and without high salt followed by incubation with goat anti-mouse HRP conjugate (1:10,000). Cos-7 cell lysates (1µg of total protein per lane) were transfected with pQM vectors encoding two different E2-tagged proteins. Detection is performed with anti-E2Tag antibody (5E11) (1:10 000 in TBST) with and without high salt followed by incubation with goat anti-mouse HRP conjugate (1:10,000).
  • Immunocytochemistry/ Immunofluorescence - Anti-E2 tag antibody [5E11] (ab977)
    Immunocytochemistry/ Immunofluorescence - Anti-E2 tag antibody [5E11] (ab977)This image is courtesy of an Abreview submitted by Dr Ashraf Mannan
    ab977 at 1/200 dilution staining human Hela cells by ICC/IF. The HeLa cells were transfected with the fusion DNA construct pQM-NTag/A consisting of E2 tag fused in frame with gene specific cDNA. The cells were fixed with paraformaldehyde and blocked with serum prior to incubation with the antibody for 2 hours. A Cy3 conjugated sheep anti-mouse IgG antibody was used as the secondary.

    See Abreview

文献

This product has been referenced in:
  • Murayama Y  et al. Establishment of DNA-DNA Interactions by the Cohesin Ring. Cell 172:465-477.e15 (2018). WB . Read more (PubMed: 29358048) »
  • Kaczmarek K  et al. Overexpression of peroxisomal testis-specific 1 protein induces germ cell apoptosis and leads to infertility in male mice. Mol Biol Cell 22:1766-79 (2011). Read more (PubMed: 21460186) »
See all 9 Publications for this product

发表研究结果有使用 ab977？请让我们知道，以便我们可以引用本数据表中的参考文章。

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Immunoprecipitation abreview for Anti-E2 tag antibody [5E11]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa)
Total protein in input
5 µg
Specification
HeLa
Immuno-precipitation step
Protein G
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Dr. Ashraf Mannan

Verified customer

提交于 Dec 22 2006

Immunocytochemistry/ Immunofluorescence abreview for Anti-E2 tag antibody [5E11]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 30%
Read More

Dr. Ashraf Mannan

Verified customer

提交于 Nov 03 2006

Question

the vial our customer got as a replacement contains only 40 µl, not 100 µ of the antibody #AB977l. What now?

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Answer

My apologies for the inconvenience, I will arrange for a new vial to be sent to you in your next shipment. Thank you for your patience,

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Question

BATCH NUMBER 9057 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No signal or weak signal. The anti-E2 antibody lot # 9057 does not or only insufficiently detect the transiently in COS7, Alexander, Hep3B and HEK 293 cells expressed fusionprotein E2-p14.5 by westernblotting This same fusionprotein is detected on the same membrane by the same antibody ordered 6 month before Lot # 6260. SAMPLE HEK 293, COS7, Alexander, Hep3B Mammalian cells PRIMARY ANTIBODY Primary Antibody Details (Dilutions, Diluting Buffer) 1.AB over night 4?C or 2hrs RT, 3x10min PBS wash SECONDARY ANTIBODY 2.AB anti mouse (Biorad) 1:3000 1hr RT 3x10 min PBS wash, DETECTION METHOD chemiluminescence by ECL POSITIVE AND NEGATIVE CONTROLS USED old Lot #6260 of the same antibody ANTIBODY STORAGE CONDITIONS The vial was always kept ? 20 C? and never touched or opened before Nov .04. Aliquots of Lot # 6260 were kept in the same box/freezer and worked fine. SAMPLE PREPARATION Lysis buffer: 20mMTRIS ph 7.4 150mMNaCl 10% Glycerol 1% TritonX100 . Complete Protein Inhibitor (Roche) 1:3000 1:5000 1:10000 in 5% skim milk 3%BSA TRANSFER AND BLOCKING CONDITIONS 30 minutes preblocking HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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Answer

I'm very sorry to hear you are having problems with ab977 lot 9057; we have not had any reported problems with this lot, it is possible the antibody has gone off when it was sent to you. I will arrange for a new vial to be sent to you as soon as possible with your shipment. We are currently out of stock of this antibody but should have some more in the next few days, my apologies for the delay. I would be very grateful if you could let me know if you are satisfied with the new lot. Thank you for your patience,

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Question

Is one of your anti-E2 tag antibodies better than the other for immunoprecipitation? How much have you used for immunoprecipitation experiments? Is one more sensitive for western blotting? If one is better for blotting and the other for immunoprecipitation, can you indicate anything about the magnitudes of the differences? Sincerely yours, John Dougherty

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Answer

For immunoprecipitation and for affinity chromatography we recommend to use ab978 - during further elution milder conditions can be used. We have used ab978 beads for affinity purification of several different proteins in small quantities. For western blot it is better to use ab977 as this clone has higher affinity than ab978.

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

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