Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)

Reviews (4)Q&A (3)References (27)

Overview

  • Product name

    Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker
    See all Cytokeratin 10 primary antibodies

  • Description

    Mouse monoclonal [DE-K10] to Cytokeratin 10 - Cytoskeleton Marker

  • Host species

    Mouse

  • Specificity

    Reacts with keratinizing stratified epithelia and in differentiated areas of highly differentiated squamous cell carcinomas.

  • Tested applications

    Suitable for: IHC-Fr, ICC, IHC-P, WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Sheep, Cat, Dog, Human, Frog

  • Immunogen

    Cytoskeletal preparation extracted from human epidermis

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Protein G purified

  • Clonality

    Monoclonal

  • Clone number

    DE-K10

  • Myeloma

    Sp2/0

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9026 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-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-P 1/100.
WB 1/100 - 1/1000.
Flow Cyt 1/100 - 1/200.

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

Target

  • Tissue specificity

    Seen in all suprabasal cell layers including stratum corneum.

  • Involvement in disease

    Defects in KRT10 are a cause of bullous congenital ichthyosiform erythroderma (BCIE) [MIM:113800]; also known as epidermolytic hyperkeratosis (EHK) or bullous erythroderma ichthyosiformis congenita of Brocq. BCIE is an autosomal dominant skin disorder characterized by widespread blistering and an ichthyotic erythroderma at birth that persist into adulthood. Histologically there is a diffuse epidermolytic degeneration in the lower spinous layer of the epidermis. Within a few weeks from birth, erythroderma and blister formation diminish and hyperkeratoses develop.
    Defects in KRT10 are a cause of ichthyosis annular epidermolytic (AEI) [MIM:607602]; also known as cyclic ichthyosis with epidermolytic hyperkeratosis. AEI is a skin disorder resembling bullous congenital ichthyosiform erythroderma. Affected individuals present with bullous ichthyosis in early childhood and hyperkeratotic lichenified plaques in the flexural areas and extensor surfaces at later ages. The feature that distinguishes AEI from BCIE is dramatic episodes of flares of annular polycyclic plaques with scale, which coalesce to involve most of the body surface and can persist for several weeks or even months.

  • Sequence similarities

    Belongs to the intermediate filament family.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • BCIE antibody
    • BIE antibody
    • CK 10 antibody
    • CK-10 antibody
    • Cytokeratin-10 antibody
    • EHK antibody
    • K10 antibody
    • K1C10_HUMAN antibody
    • Keratin 10 antibody
    • Keratin 10 type I antibody
    • Keratin antibody
    • Keratin type i cytoskeletal 10 antibody
    • Keratin type I cytoskeletal 59 kDa antibody
    • Keratin-10 antibody
    • Keratin10 antibody
    • KPP antibody
    • KRT10 antibody
    • type I cytoskeletal 10 antibody
    see all

Images

  • Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)
    Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)

    IHC image on a frozen section of  dog skin showing its strong reactivity in the keratinizing epidermal cells.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)
    IHC image of ab9026 staining in Human cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9026, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker (ab9026)Image from Reinheckel T et al, J Cell Sci. 2005 Aug 1;118(Pt 15):3387-95, Fig 3. DOI 10.1242/?jcs.02469
    Mesenchymal-epidermal interactions assessed by heterologous organotypic co-cultures (OTC). Heterologous OTC consisting of mice deficient for cathepsin L +/+ fibroblasts in collagen type I gels topped by normal human primary keratinocytes were grown air-exposed for 7 days. Paraffin sections were stained in haematoxylin and eosin (HE) or by immunohistochemistry for the proliferation marker Ki67 (Ki67, brown nuclear staining) and the differentiation markers cytokeratin 10 (K10, ab9026 1/100 dilution, brown staining) and transglutaminase (TG, brown staining). Bar 100 µm.

References

This product has been referenced in:

  • Changarathil G  et al. Wild-type and SAMP8 mice show age-dependent changes in distinct stem cell compartments of the interfollicular epidermis. PLoS One 14:e0215908 (2019). Read more (PubMed: 31091266) »
  • Lämmermann I  et al. Blocking negative effects of senescence in human skin fibroblasts with a plant extract. NPJ Aging Mech Dis 4:4 (2018). Read more (PubMed: 29675264) »
See all 27 Publications for this product

Publishing research using ab9026? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Read More

Abcam user community

Verified customer

Submitted Oct 09 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Horse Tissue sections (SKIN)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA PH9
Permeabilization
Yes - 0,05% TX 100
Specification
SKIN
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Formaldehyde
Read More

Dr. Vincenzo Miragliotta

Verified customer

Submitted Jun 05 2017

Western blot abreview for Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker

 Good
Abreviews
abreview image
Application
Western blot
Sample
Pig Tissue lysate - whole (skin)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
nurn wound with stem cell treatment
Specification
skin
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted May 20 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 10 antibody [DE-K10] - Cytoskeleton Marker

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.05% typsin
Permeabilization
No
Specification
skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Feb 23 2015

Question

Follow up to previous correspondance: Please find attached a jpeg scan of a typical blot using the antibody, (ab9026) DK-E10. This spotting pattern has been consistently seen under a variety of conditions in an effort to optimize its use. In brief: Standard protocol 5ug protein/lane PVDF membrane Block 10% skimmed milk in TBST (0.1% Tween). Primary ab 1:5000 in 1% milk/TBST. 1 hour RT I have tried a wide variety of blocking agents, and incubated the primary ab overnight at 40C. I have also tried centrifuging the primary antibody and using the supernatant with change to the pattern. Detection system I have kept standard, and works well with all other antibodies on these lysates. I would be grateful if you could resolve this problem. Alternatively might it be worth trying an alternative clone, possibly RKSE-60, (ab9025). Best wishes,

Read More
Answer

I have just received the following information from the supplier of ab9026: "1) Yes please, you can offer your customer a free (replacement) batch of our other keratin 10 ab9025. 2)The question is however, whether this will be of any help. Ab9026 seems to work perfectly well in the blotting, giving the proper mol. weight band, and I presume that in a one-dimensional SDS-gel your customer will find this one band (possible with a few fainter bands that represent the breakdown products of cytokeratin 10). In the two-dimensional gel, however, your customer finds multiple spots at the same mol. weight level. In our experience cytokeratin 10 is a difficult protein in iso-electric focussing, which I presume was used in the first dimension. You really need high (saturated; over 8M) urea in the sample buffer and in the focussing gel. Also, when using such high concentrations of urea, heating of the sample will give carbamylation of the proteins, resulting in exactly the “train” of spots as seen in your customers gel. I would therefore suggest your customer to take these items into consideration in preparing the sample and before preparing the gel/blot. " Would you like to give those suggestions a try or prefer a vial of ab9025 to be sent to you? I am away next week but my colleagues will be able to help you, please let us know if you need further assistance,

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Question

I have purchased cytokeratin 10 antibody ab9026 clone DK-E10 from you. I am attempting to probe for keratin 10 in lystates by western blotting from human oesophageal cell lines, previously detected by 2D gel electrophoresis and mass spectrometry. The antibody produces a single signal across the blot as expected at the predicted approximate molecular weight. However, the signal is a series of punctate blobs instead of a single line. The signal is also dotted around the blot. I have tried boiling and sonicating my lysate, with no improvemnet and 4 other antibodies work fine with this lysate. I have centrifuged the antibody diluted 1:500 in TBST in a microfuge at 13000rpm for ten minutes. This produces an improvement by removing the signal from around the blot. The signal around the predicted molecular weight is still in the form of punctate blobs! I would be grateful if you could provide me with a specific protocol, that will solve this problem for me, and if possible show me an image of a typical blot for this antibody. Alternatively, perhaps you could replace the antibody with another batch, or suggest an alternative clone. Best wishes,

Read More
Answer

I'm sorry to hear you are having problem with ab9026. Spotting can be frequently caused by the blocking agent you use, I would therefore try filtering it and leaving on the membrane for 30min, then rinsing the membrane with TBST. Incubate the primary antibody in TBST at 4C and rinse the membrane well, then incubate the secondary in TBST. You may also have to try different blocking agents (milk, BSA, serum). Spotting can also be caused by the secondary antibody and the detection kit, you may wish to try other secondaries and a new detection kit. Please let me know if this helps and do not hesitate to contact us if you still have problems and we will send you a replacement.

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Question

I have used your cytokeratin 10 (clone DE-K10) successfully on normal dog tissue. I would like to use your antibody for Western blots on a group of dogs containing a K10 mutation resulting in a truncated protein. Would it be possible for you to tell us specifically what aa sequence the antibody binds to so that I know if my mutated protein would be expected to bind? Thanks

Read More
Answer

The exact epitope of Ab9026 is not known. I'm afraid you just have to try and see if your truncated protein is still recognized by Ab9026.

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