概述

  • 产品名称
    Anti-DDDDK tag抗体(Agarose)
    参阅全部 DDDDK tag 一抗
  • 描述
    山羊多克隆抗体to DDDDK tag (Agarose)
  • 宿主
    Goat
  • 偶联物
    Agarose
  • 经测试应用
    适用于: IPmore details
  • 免疫原

    Full length native protein (purified) corresponding to DDDDK tag conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    xxxDDDDK

  • 常规说明
    Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with xxxDDDDK may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.

    Affinity purified antibodies were coupled to agarose beads using a cyanogen bromide method.

性能

应用

Our Abpromise guarantee covers the use of ab1240 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.

靶标

  • 相关性
    This is a useful tool for the localisation and characterisation of DDDDK tagged proteins.
  • 别名
    • DDDDK epitope tag antibody
    • DDDK antibody
    • ddk antibody
    • DYKDDDDK antibody
    • DYKDDDDK epitope tag antibody
    • DYKDDDDK tag antibody
    • ECS epitope tag antibody
    • ECS tag antibody
    • Enterokinase Cleavage Site epitope tag antibody
    • Enterokinase Cleavage Site tag antibody
    • FLAG antibody
    • FLAG tag antibody
    see all

文献

This product has been referenced in:
  • Hughes DJ  et al. NEDDylation is essential for Kaposi's sarcoma-associated herpesvirus latency and lytic reactivation and represents a novel anti-KSHV target. PLoS Pathog 11:e1004771 (2015). IP ; Human . Read more (PubMed: 25794275) »
  • Klement K  et al. Opposing ISWI- and CHD-class chromatin remodeling activities orchestrate heterochromatic DNA repair. J Cell Biol 207:717-33 (2014). Read more (PubMed: 25533843) »
See all 3 Publications for this product

客户评价及客户问答

1-8 of 8 Abreviews or Q&A

Answer

Thank you for contacting us.

If you are doing co-IP, it is not recommended to use RIPA buffer as it denaturing and may be too harsh on your protein complexes. I would suggest using a non-denaturing lysis buffer. For example:

20 mM Tris HCl pH 8
137 mM NaCl
1% Nonidet P-40 (NP-40) (Triton X-100 can be substituted for NP-40)
2 mM EDTA

Store up to 6 months at 4°C. Immediately before use add protease inhibitors.

As for your question regarding centrifugation, as shown on https://www.abcam.com/index.html?pageconfig=resource&rid=11385#d, section 4.2 "Method B", step 5, we suggest centrifuging at 1,000-3,000 g for 2 minutes at 4°C.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for your reply.

I would recommend for the first time use of using a couple of different antibody concentrations and then seeing which one gives the best signal. You could have one condition where you use 2ug of antibody and another where you use 5ug of antibody.

If there is anything else I can help you with, please let me know.

Also, since you are based in the UK and I am based on the West coast of the USA, if you wish to receive faster replies, you could email mailto:technical@abcam.com and that will go to our UK team. I am more than happy to give any assistance, I just wanted to let you know why there is a delay in my response.

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Answer

Thank you for your reply.

This product are ready to use and the total volume of each vial you receive should be 400ul (200ul of gel and 200ul of buffer). Therefore to get a 50% slurry, you would have to use 100ul of product, wash in PBS and then resuspend in 50ul of PBS, that would give you 50ul of gel and 50ul of PBS.

If there is anything else I can help you with, please let me know.

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Answer

Thank you for contacting Abcam.

You would use the product, ab1240 as you would use regular Protein A/G Sepharose beads, same type of buffers and centrifugation steps. I have attached the Abcam protocols booklet, which contains a general IP protocol that you would be able to use for guidance.

If there is anything else I can help you with, please let me know.

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Question
Answer

Vielen Dank für Ihren Anruf.

Da es für Sie so dringend ist, habe ich im Labor angerufen, und man konnte mir tatsächlich prompt Auskunft geben:

Die Konzentration beträgt 10 ug Antikoerper/200 ug "Slurry", das Volumen beträgt 400 ul.

Verwenden Sie 15 bis 25 mcl pro 0.1-1.0 mg Proteinlysate.

Ich hoffe, dies hilft Ihnen weiter.







Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Answer

Thank you for your inquiry.


Pre-clearing the lysates is always up to the end-user to determine because background levels will vary. Since both of these antibodies are conjugated to agarose, if you want to pre-clear the lysates you could use an isotype control.


For your information, we do have a suitable isotype control for both ab1233 and ab1240 as a goat IgG Agarose conjugated antibody, ab104155.


https://www.abcam.com/Goat-IgG-Agarose-ab104155.html


Also, here is our general IP protocol:


https://www.abcam.com/index.html?pageconfig=resource&rid=11385#c


I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for contacting us. I am sorry to hear you have been experiencing problems with this anti-DDDDK tag antibody. Indeed, 100 µg of the affinity purified antibody is attached to the agarose gel and the total volume of each vial you received should be 400 microliters (200 microliters of gel and 200 microliters of buffer). I am sorry that you did nor receive this for one of the vials and for the inconvenience this has caused. Therefore, I have issued a free of charge replacement with the order number 1003302 which you should receive after the christmas holidays. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

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Question
Answer

Thank you for your inquiry. Concentration: 100 ug antibody/200 ul gel Volume: 400 ul of 50% slurry containing 200 ul gel Antibody concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG, before coupling to agarose beads. I hope this information helps. Please contact us with any other questions.

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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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