Overview

  • Product name

    Anti-Cytokeratin 3/CK-3 antibody [AE5]
    See all Cytokeratin 3/CK-3 primary antibodies
  • Description

    Mouse monoclonal [AE5] to Cytokeratin 3/CK-3
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Rabbit, Cow, Human
  • Immunogen

    Tissue, cells or virus corresponding to Rabbit Cytokeratin 3/CK-3.

  • General notes

    Previously labelled as Cytokeratin 3.

Properties

Applications

Our Abpromise guarantee covers the use of ab77869 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
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB 1/1000 - 1/3000. Predicted molecular weight: 65 kDa.
Flow Cyt Use 2µg for 106 cells.

Target

  • Tissue specificity

    Cornea specific.
  • Involvement in disease

    Defects in KRT3 are a cause of Meesmann corneal dystrophy (MECD) [MIM:122100]; also abbreviated as MCD and known as juvenile epithelial corneal dystrophy of Meesmann. MECD is an autosomal dominant disease that causes fragility of the anterior corneal epithelium. Patients are usually asymptomatic until adulthood when rupture of the corneal microcysts may cause erosions, producing clinical symptoms such as photophobia, contact lens intolerance and intermittent diminution of visual acuity. Rarely, subepithelial scarring causes irregular corneal astigmatism and permanent visual impairment. Histological examination shows a disorganized and thickened epithelium with widespread cytoplasmic vacuolation and numerous small, round, debris-laden intraepithelial cysts.
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Information by UniProt
  • Database links

  • Alternative names

    • 65 kDa cytokeratin antibody
    • CK-3 antibody
    • CK3 antibody
    • Cytokeratin 3 antibody
    • Cytokeratin-3 antibody
    • K2C3_HUMAN antibody
    • K3 antibody
    • Keratin 3 antibody
    • Keratin antibody
    • Keratin type II cytoskeletal 3 antibody
    • Keratin-3 antibody
    • KRT3 antibody
    • type II cytoskeletal 3 antibody
    • Type-II keratin Kb3 antibody
    see all

Images

  • Overlay histogram showing A431 cells stained with ab77869 (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 (ab77869, 2µg/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.

  • ab77869 staining Cytokeratin 3 / CK-3 (red) in transfected Ptk2 cells by Immunocytochemistry/ Immunofluorescence.

    Cells were co-transfected in quadruplicate with wild-type, mutant, or a 1:1 mixture of wild-type and mutant K12 expression plasmids along with a wild-type K3 expression construct. After 72 hours, cells were fixed with 3% paraformaldehyde for 20 minutes, before being incubated with primary antibody. Nuclei were stained with DAPI (blue).

References

This product has been referenced in:

  • Shivakumar S  et al. Chloroquine Protects Human Corneal Epithelial Cells from Desiccation Stress Induced Inflammation without Altering the Autophagy Flux. Biomed Res Int 2018:7627329 (2018). Read more (PubMed: 30519584) »
  • Hanson C  et al. Transplanting embryonic stem cells onto damaged human corneal endothelium. World J Stem Cells 9:127-132 (2017). Read more (PubMed: 28928909) »
See all 8 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for contacting us and your interest in our products.


As discussed over the phone, the Anti-Cytokeratin 3 antibody [AE5](ab77869) should be suitable for the experiment you are hoping to perform. This particular clone has been used previously to detect corneal keratin in rabbit corneal epithelium (please refer to the reference below).


Schermer A et al. Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells. J Cell Biol 103:49-62 (1986). PubMed: 2424919


Unfortunately I have not been able to locate any additional in house data where immmunohistochemical staining of rabbit tissue has been performed. However, all of our products are covered by the Abpromise. This means that if you find the product does not work as stated on the datasheet (in this case both rabbit and paraffin embedded immunohistochemistry are guaranteed), and you report the problem within 6 months of purchase, you would be entitled to a replacement antibody or a refund.


More information on the Abpromise can be found from the following link:


https://www.abcam.com/abpromise


I hope this information has been of help. If you have any further questions please do not hesitate to ask.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (tooth)
Permeabilization
Yes - triton x-100
Specification
tooth
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 30 2013

Answer

Thank you for your inquiry and your interest in our products.

I searched our catalogue and we currently do not have a cytokeratin 3 antibody that is directly conjugated. We do carry ab77869, which has been tested in FC with rabbit, cow and human which could be conjugated to FITC using one of our EasyLink conjugation kits, like ab102885.

https://www.abcam.com/cytokeratin-3-antibody-ae5-ab77869.html

https://www.abcam.com/index.html?pageconfig=resource&rid=13148&source=pagetrap&viapagetrap=easylink

For cytokeratin 12, we do not carry any antibodies that have been tested in flow cytometry. However, if you would like to test ab124975 or ab113039 in FC, we could offer you a discount on a future purchase if you purchase, test, and submit an Abreview on the product (the discount is worth the price you pay for the antibody).

For p63, we have an antibody conjugated to DyLight 488 that works in ICC/IF with human cells, ab139929. Again, if you would like to test this antibody in FC, we could offer you a future discount. https://www.abcam.com/p63-antibody-epr5701-dylight-488-ab139929.html

For BCRP/ABCG2, we have 4 antibodies tested in FC, but non are conjugated to a fluorophore: ab24115, ab24114, and ab72788, and ab95692 (conjugated to biotin). You could use the purified antibody ab72788 with one of our conjugation kits to directly conjugate a fluorophore to the antibody.

I hope this information helps. If you are interested in testing any of the antibodies as listed above, please let me know and I will set you up in our system.

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Answer

Thank you for contacting Abcam.

I will be happy to help you further in this matter, but I just have a few more questions for you:

1 - Were you staining tissue sections or a monolayer of cultured cells?

2 - If you were staining sections, how were they prepared (PfA fixation & paraffin embedded, MeOH etc)?

3 - If these samples were paraffin embedded, what type of antigen retrieval did you use on them?

4 - Have you used either of the secondary antibodies successfully with other primary antibodies?

I look forward to your reply and helping to resolve these issues.

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Answer

Thats no problem. If you have any further questions please do not hesitate to contact us again.

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Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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