Recombinant
RabMAb

Recombinant Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)

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Overview

  • Product name

    Anti-Keratin 12/K12 antibody [EPR17882]
    See all Keratin 12/K12 primary antibodies

  • Description

    Rabbit monoclonal [EPR17882] to Keratin 12/K12

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Keratin 12/K12 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q99456

  • Positive control

    • WB: Keratin 12/K12 transfected HEK-293 whole cell lysate; Mouse and rat eyeball lysates. IHC-P: Mouse and rat cornea tissues. ICC/IF: Keratin 12/K12 transfected HeLa cells.

  • General notes

     

     

    Previously labelled as Keratin 12. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab185627 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 1/50.
WB 1/5000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested in mouse and rat.

Target

  • Function

    May play a unique role in maintaining the normal corneal epithelial function. Together with KRT3, essential for the maintenance of corneal epithelium integrity.

  • Tissue specificity

    Cornea specific.

  • Involvement in disease

    Defects in KRT12 are a cause of Meesmann corneal dystrophy (MECD) [MIM:122100]; also abbreviated 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

    see all

  • Alternative names

    • CK-12 antibody
    • Cytokeratin-12 antibody
    • K12 antibody
    • K1C12_HUMAN antibody
    • Keratin 12 antibody
    • Keratin antibody
    • Keratin, type I cytoskeletal 12 antibody
    • Keratin-12 antibody
    • KRT12 antibody
    • type I cytoskeletal 12 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)

    Immunohistochemical analysis of paraffin-embedded Mouse cornea tissue labeling Keratin 12/K12 with ab185627 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on the mouse corneal epithelium is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)

    Immunohistochemical analysis of paraffin-embedded Rat cornea tissue labeling Keratin 12/K12 with ab185627 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on the rat corneal epithelium is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    Immunocytochemistry/ Immunofluorescence - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Keratin 12/K12 with ab185627 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing no staining on HeLa cell line.
    The expression increased after transfection of Keratin 12/K12. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab185627 at 1/50 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Western blot - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    Western blot - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    All lanes : Anti-Keratin 12/K12 antibody [EPR17882] (ab185627) at 1/100000 dilution

    Lane 1 : Empty vector (vector control) transfected HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 2 : Keratin 12/K12 transfected HEK-293 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 54 kDa
    Observed band size: 54 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    Western blot - Anti-Keratin 12/K12 antibody [EPR17882] (ab185627)
    All lanes : Anti-Keratin 12/K12 antibody [EPR17882] (ab185627) at 1/5000 dilution

    Lane 1 : Mouse eyeball lysate
    Lane 2 : Rat eyeball lysate
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 4 : Y79 (Human retinoblastoma cell line) whole cell lysate
    Lane 5 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 7 : Rat brain lysate
    Lane 8 : Rat heart lysate
    Lane 9 : Rat kidney lysate
    Lane 10 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 54 kDa
    Observed band size: 54 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Keratin 12/K12 is expressed specifically in corneal epithelium (Am J Hum Genet. 1997. 61, 1268–1275).

References

This product has been referenced in:

  • Zhang R  et al. Mechanical strain triggers differentiation of dental mesenchymal stem cells by activating osteogenesis-specific biomarkers expression. Am J Transl Res 11:233-244 (2019). Read more (PubMed: 30787982) »
  • Hu L  et al. Expansion and maintenance of primary corneal epithelial stem/progenitor cells by inhibition of TGFß receptor I-mediated signaling. Exp Eye Res 182:44-56 (2019). Read more (PubMed: 30914160) »
See all 8 Publications for this product

Publishing research using ab185627? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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IHC - Wholemount abreview for Anti-Keratin 12/K12 antibody [EPR17882]

 Average
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (cornea)
Specification
cornea
Read More

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Verified customer

Submitted May 17 2019

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