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Anti-CHD8 antibody (ab114126)

Reviews (1)Q&A (5)References (6)
Western blot - Anti-CHD8 antibody (ab114126)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD8 antibody (ab114126)
  • Immunoprecipitation - Anti-CHD8 antibody (ab114126)
  • Western blot - Anti-CHD8 antibody (ab114126)

Key features and details

  • Rabbit polyclonal to CHD8
  • Suitable for: WB, IP, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-CHD8 antibody
    See all CHD8 primary antibodies

  • Description

    Rabbit polyclonal to CHD8

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rabbit, Guinea pig, Chimpanzee, Rhesus monkey, Gorilla, Chinese hamster, Orangutan, Elephant

  • Immunogen

    Synthetic peptide, corresponding to a region within amino acids 325-350 of Human CHD8 (NP_065971.2)

  • Positive control

  • General notes

    Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.

    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

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • Storage buffer

    pH: 7
    Preservative: 0.09% Sodium azide
    Constituent: 99.91% Tris citrate/phosphate

    pH: 7-8
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Purification notes

    ab114126 was affinity purified using an epitope specific to CHD8 immobilized on a solid support.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab114126 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
WB
1/2000 - 1/10000. Predicted molecular weight: 291 kDa.
IP
Use at 2-5 µg/mg of lysate.
IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/2000 - 1/10000. Predicted molecular weight: 291 kDa.
IP
Use at 2-5 µg/mg of lysate.
IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    DNA helicase that acts as a chromatin remodeling factor and regulates transcription. Acts as a transcription repressor by remodeling chromatin structure and recruiting histone H1 to target genes. Suppresses p53/TP53-mediated apoptosis by recruiting histone H1 and preventing p53/TP53 transactivation activity. Acts as a negative regulator of Wnt signaling pathway by regulating beta-catenin (CTNNB1) activity. Negatively regulates CTNNB1-targeted gene expression by being recruited specifically to the promoter regions of several CTNNB1 responsive genes. Involved in both enhancer blocking and epigenetic remodeling at chromatin boundary via its interaction with CTCF. Acts as a suppressor of STAT3 activity by suppressing the LIF-induced STAT3 transcriptional activity. Also acts as a transcription activator via its interaction with ZNF143 by participating to efficient U6 RNA polymerase III transcription.

  • Sequence similarities

    Belongs to the SNF2/RAD54 helicase family. CHD8 subfamily.
    Contains 2 chromo domains.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.

  • Post-translational
    modifications

    Sumoylated.

  • Cellular localization

    Nucleus. Localizes to the promoter regions of several CTNNB1-responsive genes. Also present at known CTCF target sites.
  • Information by UniProt

  • Database links

    • Alternative names

      • ATP dependent helicase CHD8 antibody
      • ATP-dependent helicase CHD8 antibody
      • Axis duplication inhibitor antibody
      • Beta catenin binding protein like antibody
      • CHD 8 antibody
      • CHD-8 antibody
      • chd8 antibody
      • CHD8_HUMAN antibody
      • Chromodomain helicase DNA binding protein 8 antibody
      • Chromodomain-helicase-DNA-binding protein 8 antibody
      • DKFZp686N17164 antibody
      • Duplin antibody
      • Helicase with SNF2 domain 1 antibody
      • HELSNF1 antibody
      • KIAA1564 antibody
      see all

    Images

    • Western blot - Anti-CHD8 antibody (ab114126)
      Western blot - Anti-CHD8 antibody (ab114126)
      All lanes : Anti-CHD8 antibody (ab114126) at 0.1 µg/ml

      Lane 1 : HeLa whole cell lysate at 50 µg
      Lane 2 : HeLa whole cell lysate at 15 µg
      Lane 3 : HeLa whole cell lysate at 5 µg

      Developed using the ECL technique.

      Predicted band size: 291 kDa


      Exposure time: 30 seconds
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD8 antibody (ab114126)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD8 antibody (ab114126)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling CHD8 with ab114126 at 1/1000 (1µg/ml). Detection: DAB.
    • Immunoprecipitation - Anti-CHD8 antibody (ab114126)
      Immunoprecipitation - Anti-CHD8 antibody (ab114126)
      ab114126 at 3ug/mg of lysate detecting CHD8 by immunoprecipitation (lane 1). 1mg of HeLa whole cell lysate was used, 20% of immunoprecipitate was loaded. Lane 2: IgG control.
      Detection: chemiluminescence. Exposure time: 30 seconds.
    • Western blot - Anti-CHD8 antibody (ab114126)
      Western blot - Anti-CHD8 antibody (ab114126)
      All lanes : Anti-CHD8 antibody (ab114126) at 0.1 µg/ml

      Lane 1 : HeLa whole cell lysate
      Lane 2 : 293T whole cell lysate
      Lane 3 : Jurkat whole cell lysate
      Lane 4 : TCMK-1 whole cell lysate

      Lysates/proteins at 50 µg per lane.

      Developed using the ECL technique.

      Predicted band size: 291 kDa


      Exposure time: 3 minutes

    References (6)

    Publishing research using ab114126? Please let us know so that we can cite the reference in this datasheet.

    ab114126 has been referenced in 6 publications.

    • Fan X  et al. TWIST1 and chromatin regulatory proteins interact to guide neural crest cell differentiation. Elife 10:N/A (2021). PubMed: 33554859
    • Lee CY  et al. A spontaneous missense mutation in the chromodomain helicase DNA-binding protein 8 (CHD8) gene: a novel association with congenital myasthenic syndrome. Neuropathol Appl Neurobiol 46:588-601 (2020). PubMed: 32267004
    • Zhao C  et al. Dual Requirement of CHD8 for Chromatin Landscape Establishment and Histone Methyltransferase Recruitment to Promote CNS Myelination and Repair. Dev Cell 45:753-768.e8 (2018). PubMed: 29920279
    • Wade AA  et al. Common CHD8 Genomic Targets Contrast With Model-Specific Transcriptional Impacts of CHD8 Haploinsufficiency. Front Mol Neurosci 11:481 (2018). PubMed: 30692911
    • Gompers AL  et al. Germline Chd8 haploinsufficiency alters brain development in mouse. Nat Neurosci 20:1062-1073 (2017). WB, ChIP-sequencing ; Mouse . PubMed: 28671691
    • Cotney J  et al. The autism-associated chromatin modifier CHD8 regulates other autism risk genes during human neurodevelopment. Nat Commun 6:6404 (2015). IP, ChIP . PubMed: 25752243

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-6 of 6 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-CHD8 antibody

     Poor
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Foregut)
    Permeabilization
    No
    Specification
    Foregut
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
    Fixative
    4% 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

    Submitted Mar 14 2019

    Question

    I can wait for that. Thanks.

    Read More
    Answer

    Thank you for your reply.

    I have gone ahead and processed the free of charge replacement, your new order number is * and we will ship the antibody to you as soon as we can.

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

    Read More

    Question

    Thank you for a prompt solution. I look forward to trying out the new antibody.

    Read More
    Answer

    Thank you for your reply.

    I just realized whilst I was trying to process the replacement that the ab57553, that it is currently out of stock, and we have an estimated delivery date of 24 July. Is this OK with you? I wanted to check in and ask you first before I finalize the replacement order. The alternative is that I could refund the antibody.

    I look forward to your reply.

    Read More

    Question

    Thank you for getting back to me promptly. Here are the answers for the additional questions.





    Read More
    Answer

    Thank you for your reply and for providing that information.

    I am sorry that the antibody was not working for you and based on the information that you have provided, I think the best course of action, is for me to either replace the antibody for you, or to process a full refund.

    One possible antibody that I could send as a replacement is, ab27553:

    https://www.abcam.com/CHD8-antibody-ab57553.html



    Please let me know how you would like to proceed and I will be happy to help resolve this issue for you.

    I look forward to your reply.

    Read More

    Question

    First batch worked on a couple of Western blots and stopped working. Initially I thought the antibody got degraded by protease, etc. so I ordered another batch and never got Western blot working ever since. I'd like a replacement for another product (ab57553). Amount of protein loaded: 10ug HEK lysate Blocking: 10% dry milk + 5% normal goat serum in TBST Detection method: ECL and Licor infrared 7.5% gel ran at 130V, transferred at 80V 400mA 5 hours Primary antibody conditions: 1:1000, 1:500, 1:250 for 2hr up to o/n Secondary: rabbit HRP from Amersham 1:10000, 1hr RT washing 3 times in TBST

    Read More
    Answer

    Thank you for contacting Abcam.

    I am sorry about the issues you are having with ab114126 in western blot. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

    I would just like to ask a few more questions:

    1 - Can I ask how the antibody was being stored? Was if frozen at -20C or kept at 4C?

    2 - You block in 10% dry milk + 5% normal goat serum, have you tried just using 5% milk? I ask as the blocking agent you are using is different than what is normally used.

    3 - When you did get a signal using the antibody, was it at the correct/expected molecular weight?

    Also, would you be able to provide me with either the Abcam order number or PO# that was used in the purchase of the antibody,that would be very helpful.

    I look forward to your reply.

    Read More

    Question

    What is the definition of ChIP grade?

    Read More
    Answer

    Thank you for calling Abcam. I just talked to a colleague and they informed me that when an antibody had been labeled ChIP grade, it means that Abcams in-house ChIP expert has reviewed all of the available ChIP data on the antibody and determined that is it suitable for the application. We have many antibodies in our catalogue that have been successfully used in ChIP assays, however the volume of data on them remains relatively low and so we have not yet classed them as ChIP grade. If there is anything else i can help with, please let me know.  

    Read More

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