Overview

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 0.5% BSA, Tris buffered saline, pH 7.3
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Fast track antibodies constitute a diverse group of products that have been released to accelerate your research, but are not yet fully characterized. They have all been affinity purified and show high titre values against the immunizing peptide (by ELISA). Fast track terms of use

Application Abreviews Notes
WB
ELISA
  • Application notes
    Peptide ELISA: Use at an assay dependent dilution. Antibody detection limit dilution: 1/128000.
    WB: Use at an assay dependent dilution. Predicted molecular weight: 63 kDa.
    Preliminary experiments gave an approx 100kDa band in Human Brain (Cerebellum) lysates after 0.2µg/ml antibody staining. The 100kDa band was successfully blocked by incubation with the immunizing peptide.
    Please note that currently we cannot find an explanation in the literature for the size of the band we observe.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    References

    ab77432 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Question

    And my purpose is to detect the expression of TOX3 in breast cancer cell lines and tissues with immunohistochemistry and Westernblot. I have bought some Goat anti-TOX3(ab77432) antibodies from your company.    First my colleague and me tried to detect the expression of TOX3 in 12 normal breast cell lines and breast cancer cell lines with westernblot. As your datasheet shows, we found one band is around 100kDa. Then  I was going to stain TOX3 with the same antibody in huam tissues. So I chose normal huamn brain tissue as positive control. After the staining with immunohistochemistry, I found clear positive signal in the cytoplasm of the neuron cells.But it says that TOX3 protein possibly localized  in the nuclei. Althoung I haven't finished the optimisation of the delution of the antibody until now,  here I can not see any signal in the nuclei.  my immunohistochemistry protocal is as follows: antigen retrieval: EDTA-Tris pH9.0, Microwave blocking for peroxidase blocking for endogenouse biotin with the Avidin-Biotin Kit from Invitrogen blocking non-specifical staining with 10% normal rabbit serum anti-TOX3(ab77432) 1:50 room temperature(RT) 1 hour; PBS as negative control rabbit anti-goat IgG (biotin conjugated, DAKO) 1:500 in RT 3Omin HRP in RT 15min DAB hematoxylin for counter staining   So my question is that if you have received such kind of feedback or do you have detected the specificity of this antibody? Could you give me some suggestion to my research with this antibody?   And I would like to send some pictures of my staining with your antibody.  I hope it will be more helpful.   Looking forward to hearing from you soon! Thanks a lot!  

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    Answer

    Thank you for your enquiry and for including the IHC and western blot images. It does seem the antibody is working as expected in western blotting. Until now we have not had any feedback regarding the use of ab77432 in IHC. I agree that you are seeing cytoplasmic staining in your sections and that, since TOX3 is a transcription factor it should be localized to the nucleus. It may be that this antibody is not useful for IHC. However it might be worth trying different antigen retrieval. For example, you could try using a pH6 buffer for heat induced retrieval. Alternatively, you could try a protease treatment. I hope this is helpful. Please contact me again if you have any further questions.

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