Overview

  • Product name
    Anti-Mucin 5AC antibody [1-13M1]
    See all Mucin 5AC primary antibodies
  • Description
    Mouse monoclonal [1-13M1] to Mucin 5AC
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IHC-P, ELISA, WB, ICCmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Mucin isolated from an ovarian cyst fluid (pure endocervical type according to the Fenoglio's classification) (Human).

  • Epitope
    The epitope is destroyed by beta-mercaptoethanol.
  • Positive control
    • Human intestine and stomach

Properties

Applications

Our Abpromise guarantee covers the use of ab24070 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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
IHC-P 1/100.
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 130 kDa.
ICC Use at an assay dependent concentration.

Target

  • Relevance
    Mucins are high molecular weight glycoproteins with 80% carbohydrates and 20% core protein. Gastric Mucin 5AC antigen is found in columnar mucus cells of surface gastric epithelium and in goblet cells of the fetal and precancerous colon but not in normal colon. Resurgence of gastric mucin during colonic carcinogenesis is suggestive of either re-expression of the peptide core of gastric mucin in the adult colon or due to changes in the glycosylation pattern of mucin, which expose the hidden Mucin 5AC antigen.
  • Cellular localization
    Secreted; Cytoplasmic and cell surface
  • Database links
  • Alternative names
    • Apomucin antibody
    • Apomucin Major Airway Glycoprotein antibody
    • Gastric mucin antibody
    • LeB antibody
    • Lewis B blood group antigen antibody
    • Major Airway Glycoprotein antibody
    • MUC 5 antibody
    • MUC5 antibody
    • MUC5AC antibody
    • Mucin 5 subtype AC tracheobronchial antibody
    • Mucin 5 Subtypes A And C antibody
    • Mucin 5 subtypes A and C tracheobronchial/gastric antibody
    • Mucin 5, tracheobronchial/gastric antibody
    • Mucin 5AC antibody
    • Mucin 5AC oligomeric mucus/gel forming antibody
    • Mucin 5AC, oligomeric mucus/gel forming pseudogene antibody
    • Mucin antibody
    • TBM antibody
    • Tracheobronchial Mucin (TBM) antibody
    • Tracheobronchial mucin antibody
    see all

Images

  • Overlay histogram showing A549 cells stained with ab24070 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24070, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Stinson SE  et al. D prostanoid receptor 2 (chemoattractant receptor-homologous molecule expressed on TH2 cells) protein expression in asthmatic patients and its effects on bronchial epithelial cells. J Allergy Clin Immunol 135:395-406 (2015). Read more (PubMed: 25312757) »
  • Wan LY  et al. Modulation of Mucin mRNA (MUC5AC and MUC5B) Expression and Protein Production and Secretion in Caco-2/HT29-MTX Co-cultures Following Exposure to Individual and Combined Fusarium Mycotoxins. Toxicol Sci 139:83-98 (2014). Read more (PubMed: 24496642) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Answer

Thank you for your phone call. I did some research regarding PVA and found that for unknown reasons, PVA does not seem to effectively block background in WB. You may need to switch to BSA or milk in order to get clear bands in addition to the new secondary I am sending.
As requested, I have issued a free of charge replacement with the order number 1203530.

Regarding ab24070, the datasheet does indicate that B mercaptoethanol destroys the epitope, so you will need to be careful to avoid this reagent when blotting for Mucin 5AC.

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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Answer

Thank you for contacting us. Looking at your blots I think there is a few things you can try. One is to decrease the secondary antibody dilution. Since the blots are all the same it indicates some nonspecific binding. You can also add a no primary control to see if there is non-sepcific binding from the secondary alone. You can also increase your blocking time (even overnight at 4 degrees) or switch your blocking agent from milk to BSA or vice versa to see if this reduces background. Normally the primary antibody dilution for a WB when using a purified antibody can be about 1 ug/ml, however since the pattern is the same for all three different antibodies I am not convinced this is a primary antibody problem. Please let me know if these suggestions don't improve your results.

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Question
Answer

Thank you for your call yesterday and for your patience while I have been in touch with the lab.

I've attached an article in which both of these Mucin 5AC antibodies were tested in Western blot. Please see page 191 for the images.

I hope this information will be useful, but if you have any further questions please let me know and I'll be happy to help.

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Answer

Thank you for your enquiry.

The immunogen used to produce ab24070 is Mucin isolated from human ovarian cyst fluid (pure endocervical type according to the Fenoglio's classification). The epitope ab24070 reacts with has not been mapped so unfortunately there are no further details available regarding the sequence specificity of this antibody.

I hope this is helpful. Please contact me again if you have any further questions.

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Answer

Thanks for sending the additional protocol information. I am sending a replacement free of charge vial of ab24070 on the order ***, which should arrive tomorrow. Please keep me updated on the stability of this antibody, and let me know if there is anything else I can help with.

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Answer

Thanks for your call today and for sending the information. I agree that the aliquot from August is not working nearly as well as expected. Could you also send a brief overview of your protocol and the storage conditions, for our documentation of the problem? Do you have your order number? I would be happy to send a replacement vial to you if you would like. Please let me know if you have any questions. I look forward to hearing from you.

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Question
Answer

Thank you for contacting us. We recommend storing this antibody at -20°C or -80°C long-term. This antibody will be viable for at least a year when frozen, but repeated freezing and thawing may damage the antibody. Frost-free freezers prevent the build up of frost by cycling between freezing and thawing. Frosted freezers are highly recommended for the storage of antibodies. I hope this helps, please let me know if you need any additional information or assistance.

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Question
Answer

Thank you for your enquiry. Following correspondence with the lab I can tell you that intestine and stomach were used as positive controls for the testing of this antibody. If you have any additional questions please let me know.

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Answer

Thank you for your enquiry. The originator of the antibody stated the following: Ethanol Fixed tissues are imbedded into paraffin in some countries. But I understand that the datasheet is unclear. We will delete the ethanol fixation with paraffin from our information. For paraffin sections: Protease pretreatment is recommended. If you have any additional questions or remarks, please do not hesitate to contact us immediately.

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Answer

Thank you for getting back to me. We currently stock several human intestine and stomach lysates. For example: ab29276 Small Intestine (Human) Whole Cell Lysate - adult normal tissue and; ab29681 Stomach (Human) Whole Cell Lysate - adult normal tissue For further information I would advise you search using the terms "Intestine" and "Stomach". I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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1-10 of 11 Abreviews or Q&A

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

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