Overview

  • Product name

    Anti-HB9/HLXB9/MNX1 antibody
    See all HB9/HLXB9/MNX1 primary antibodies
  • Description

    Sheep polyclonal to HB9/HLXB9/MNX1
  • Host species

    Sheep
  • Specificity

    ab59795 is specific for for HB9/HLXB9/MNX1.

  • Tested applications

    Suitable for: IHC-P, WB, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human HB9/HLXB9/MNX1 aa 68-88.
    Sequence:

    H-APADRLRAESPSPPRLLAAHC-NH2

  • Positive control

    • Normal Term and First Trimester human Placenta and mouse Neuroblastoma and Spinal Cord cells.
  • General notes

    For additional stability glycerol at a concentration of 1:1 can be added.

     This product was previously labelled as HB9/HLXB9

     

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab59795 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
IHC-P Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 41 kDa.
ELISA 1/500 - 1/6500.

Target

  • Function

    Putative transcription factor involved in pancreas development and function.
  • Tissue specificity

    Expressed in lymphoid and pancreatic tissues.
  • Involvement in disease

    Defects in MNX1 are a cause of Currarino syndrome (CURRAS) [MIM:176450]. The triad of a presacral tumor, sacral agenesis and anorectal malformation constitutes the Currarino syndrome which is caused by dorsal-ventral patterning defects during embryonic development. The syndrome occurs in the majority of patients as an autosomal dominant trait.
  • Sequence similarities

    Contains 1 homeobox DNA-binding domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • HB 9 antibody
    • HB9 antibody
    • HLXB 9 antibody
    • HLXB9 antibody
    • Homeo box HB9 antibody
    • Homeobox HB9 antibody
    • Homeobox protein HB9 antibody
    • HOXHB9 antibody
    • MNX1 antibody
    • MNX1_HUMAN antibody
    • Motor neuron and pancreas homeobox 1 antibody
    • Motor neuron and pancreas homeobox protein 1 antibody
    • Sacral agenesis autosomal dominant (Currarino triad) antibody
    • SCRA 1 antibody
    • SCRA1 antibody
    see all

References

This product has been referenced in:

  • Zoldan J  et al. The influence of scaffold elasticity on germ layer specification of human embryonic stem cells. Biomaterials 32:9612-21 (2011). IHC-P ; Human . Read more (PubMed: 21963156) »
  • Neufing PJ  et al. Expression and localization of homeodomain proteins DLX4/HB9 in normal and malignant human breast tissues. Anticancer Res 23:1479-88 (2003). IHC-P ; Human . Read more (PubMed: 12820413) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

I have found a paper describing development of the HB9 antibody ab59795 and the protocol that was used for IHC on tumor tissue.

Neufing PJ et al. Expression and localization of homeodomain proteins DLX4/HB9 in normal and malignant human breast tissues. Anticancer Res 23:1479-88 (2003). PubMed: 12820413

http://us.adminsite.com/admin/abcamshare/CRM/attach/2012/1/16/CCE3425883_anticancerresearch23.pdf

What I think is important to note is that the tissues were fixed with methacarn (60% methanol, 30% chloroform, and 10% glacieal acetic acid), and not a formaldehyde-based fixative, and that they did not use antigen retrieval. I realize this does not help you, since your tissues are already fixed and embedded, but a retrieval that does not use heat (e.g. enzymatic retrieval with trypsin) may be worth trying, as heat-mediated retrieval can lead to the non-specific staining you see.

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Answer

I will send the IHC protocol for ab59795 when I receive it, hoepfully in the next day or two. As I mentioned, Abcam does not validate this antibody for IHC in-house. We do not have free samples of any of our antibodies, but ab59795 is covered by our Abpromise guarantee for this application (IHC on paraffin-embedded tissue sections) and this species (human). If I cannot help you obtain a satisfactory result, I will be happy to look into replacing this. However, I am concerned about the possility that this transcription factor is not present in prostate. Do you have any other tissues you can include as apositive control, in particular pancreas?

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Answer

Thank you for sending the details of your protocol. I agree that the staining you see for both of these antibodies is not what is expected. I think what you want is shown on the datasheet for a different antibody against HB9, ab92606, showing a stain of mouse pancreas: https://www.abcam.com/HB9-HLXB9-antibody-ab92606.html As you mention, the two concerns are the backgound staining of the cytoplasm and stroma, and the lack of nuclear stain. The background may be due to the secondary and/or substrate. The no-primary negative control I mentioned should indicate if that might be the case. Is the secondary from Vector biotinylated? If so, you may need a block for endogenous biotin, as well as a block for endogenous peroxidase, though this does not appear to be an issue with the other antibody you mentioned, "ab0000". (Do you know the catalogue number for this?) The lack of a nuclear stain may reflect low expression in prostate tissue. Pancreas is a suitable positive control. I have asked the laboratory that validated the antibody for IHC for their antigen retrieval recommendation, which we do not have on file. It may be that heat-mediated AR is inappropriate. If you do try another run, I recommend including a section that does not receive any antigen retrieval. Finally, the notes on the datasheet indicate that the antibody was validated on tissue that was fixed with Methacarn, which does not include formaldehyde. (Methacarn: 60% methanol, 30% Chloroform and 10% glacial acetic acid). This suggests that no antigen retrieval was performed. The antibody may be incompatible with formalin or PFA fixation, but a different AR approach, for instance an enzymatic retrieval, may give a better result. We do not, however, have any data demonstrating this. I will get more information from the laboratory regarding the IHC validation and will send this on to you. If you have any questions or concerns, please contact me.

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Answer

Thank you for bringing this to our attention. Can you please send a few more details of your protocol? In particular, I would like to know if you included an antigen retrieval step. Also, did you confirm that the secondary is not the issue, by leaving out the primary in a negative control? What concentraion or dilution of antibody (ab59795) did you apply to the sections, and for how long? If you can attach an image of one of the stained sections, that will help. I llok forward to your reply.

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