Overview

  • Product name
    Anti-Mucin 5AC antibody [45M1]
    See all Mucin 5AC primary antibodies
  • Description
    Mouse monoclonal [45M1] to Mucin 5AC
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Tissue, cells or virus corresponding to Mucin 5AC. Prepared from the fluid of human ovarian mucinous cyst.

  • Positive control
    • IHC-P: Mouse and human Stomach and Human colon. FC: A549 cells
  • General notes

    This product was changed from ascites to tissue culture supernatant on 20th November 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab3649 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 0.5-1µg for 106 cells.

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

Use at 1 - 2 µg for 106 cells for ascites version.

IHC-P Use a concentration of 1 - 2 µg/ml.

Use at 1/100 dilution for Ascites version

ICC/IF Use a concentration of 1 - 2 µg/ml.

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

  • ab3649 staining stomach by IHC-P.
  • Overlay histogram showing A549 cells stained with ab3649 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and 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 (ab3649, 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. This antibody gave a positive signal in A549 cells fixed with 80% methanol (5 min) used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • Immunohistochemical analysis of rat stomach tissue, staining Mucin 5AC with ab3649.

    Antigen retrieval was performed via heat mediation in a citrate buffer. Samples were blocked before incubating with primary antibody (1/100) for 1 hour at room temperature. An HRP-conjugated anti-mouse IgG was used as the secondary antibody and staining was detected using DAB.
  • Immunohistochemical analysis of human stomach tissue, staining Mucin 5AC with ab3649.

  • Immunohistochemical analysis of human stomach Adenocarcinoma, staining Mucin 5AC with ab3649

  • Immunohistochemical analysis of human colon Adenocarcinoma, staining Mucin 5AC with ab3649.

References

This product has been referenced in:
  • Kohanski MA  et al. Solitary chemosensory cells are a primary epithelial source of IL-25 in patients with chronic rhinosinusitis with nasal polyps. J Allergy Clin Immunol N/A:N/A (2018). Read more (PubMed: 29778504) »
  • Guo D  et al. Ocular surface pathogenesis associated with precocious eyelid opening and necrotic autologous tissue in mouse with disruption of Prickle 1 gene. Exp Eye Res 180:208-225 (2018). Read more (PubMed: 30590023) »
See all 39 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH8
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Aug 26 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (stomach)
Permeabilization
No
Specification
stomach
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Aug 22 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (stomach)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH8
Permeabilization
No
Specification
stomach
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% buffered normal formalin

Abcam user community

Verified customer

Submitted Aug 22 2016

Answer

Unfortunately, the specific epitope of ab3649 has not been mapped to our knowledge, so we do not know where within the Mucin 5AC protein the antibody binds.

Read More

Question

As requested we send this form required for the product replacement.

Abcam IHC questionnaire



1) Abcam product code ab 3649



2) Abcam order reference number or product batch number LOT# GR101525



3) Description of the problem the staining did not work or it was extremely weak even at high concentration



4) Sample preparation:

Species: Mouse

Type of sample: PFA/formalin fixed paraffin embedded sections

Sample preparation: formalin fixed paraffin embedded sections

Positive control: mouse stomach

Negative control: mouse heart



5) Fixation step No



6) Antigen retrieval method: none or heat in citric buffer



7) Permeabilization method: No


8) Blocking agent: Donkey serum 10%

Blocking time: 20 min

Blocking temperature: RT



9) Endogenous peroxidases blocked? Endogenous biotins blocked? No



10) Primary antibody:

Concentration or dilution: 1:100 as suggested and then we increased at 1:10

Diluent buffer PBS

Incubation time 1h or over night



11) Secondary antibody:

Species: donkey

Reacts against: mouse IgG

Concentration or dilution 1:100

Diluent buffer PBS

Incubation time 1h

Fluorochrome or enzyme conjugate FITC or TRITC



12) Washing after primary and secondary antibodies:

Buffer PBS

Number of washes 5



13) Detection method Fluorescence and confocal microscope



14) How many times have you run this staining? 5 times

Do you obtain the same results every time? Yes

What steps have you altered to try and optimize the use of this antibody? Antigen retrieval method and\or Primary antibody concentration

Read More
Answer

Thank you for getting back to me and for providing some experimental details.

The protocol looks fine to me. It may well be that the antibody has been damaged during storage or shipment.

This antibody has not been tested with Mouse in IHC-P in the Lab but based on these 3 specification, it is suitable for immonohistochemistry on murine samples.

1. Wallace JL et al. Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair. Am J Pathol 178:1126-33 (2011). IHC-P; Mouse. PubMed: 21356364

2. Chen G et al. Foxa2 programs Th2 cell-mediated innate immunity in the developing lung. J Immunol 184:6133-41 (2010). IHC-P; Mouse.

3. Chen G et al. SPDEF is required for mouse pulmonary goblet cell differentiation and regulates a network of genes associated with mucus production. J Clin Invest 119:2914-24 (2009). IHC-P; Mouse. PubMed: 19759516

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

Would you be so kind to confirm the Abcam Order Number or the PON? This information is necessary for me to have to proceed with your enquiry.

I look forward to hearing from you soon.

Read More

Answer

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung)
Specification
Lung
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted May 15 2012

Answer

Thank you for your reply.

To clarify, the discount I offered is not a free sample, but can be used to get a free antibody on a future order after submitting an Abreview for ab79082.

This antibody was validated using formalin-fixed paraffin embedded tissue, or formaldehyde-fixed frozen tissue. In IHC-P, antigen retrieval was performed using citrate buffer, pH 6 in the microwave. Sections were blocked using BSA or normal serum from the host species of the secondary antibody. The antibody was diluted 1:50 - 1:100 in PBS and incubated for 1 hour at room temperature or overnight at 4C.

We would expect ab3649 and ab79082 to give the same results in all applications and speciesbecause they are the same clone. In very rare instances, the conjugated forms will behave differently, which is why we do not automatically add all of the applications and speciesto every datasheet for the same clone. The tested applications and specieslisted on our datasheets reflect what hasbeen tested using that particular format.

I hope this helps, please let me know if you need any additional information or assistance.

Read More

Answer

Yes, Sapphire Bioscienceis our distributor in Australia.

DISCOUNT CODE: ***
Expiration date: August 28th, 2012

I am very pleased to hear you would like to accept our offer and test ab79082 in canine. This code will give you: 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview forcanine and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for contacting us.

Ourantibodies to Mucin 5AC that have been validated for use on frozen tissues are primarily mouse monoclonal antibodies raised against the full length protein, for which the epitopes have not been mapped. Additionally, I have been unable to locate a full length canine Mucin 5AC sequence that I can compare to our immunogen sequences. Unfortunately, I am unable to check the homologies for these reasons.

Of the antibodies we currently sell, I would recommend the 45M1 clone: ab3649 or ab79082 (biotinylated). This clone has been shown to react with mouse, rat, rabbit, chicken, cat, human, pig, monkey, and hedgehog. Because of its broad species reactivity, it would likely also react with dog samples.

If you would like to test this antibody, please let me know, and I will be happy to offer you our 100% testing discount program:

https://www.abcam.com/collaborationdiscount

I hope this helps, please let me know if you have any further questions or concerns.

Read More

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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