Anti-pan Keratin antibody [80] (ab8068)

Reviews (8)Q&A (10)References (22)

Overview

  • Product name

    Anti-pan Keratin antibody [80]
    See all pan Keratin primary antibodies

  • Description

    Mouse monoclonal [80] to pan Keratin

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, IHC-Fr, IHC-P, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Callus cytokeratins isolated from fresh human skin tissue

  • Positive control

    • Human callus or keratinocytes

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: 0.07% Sodium azide
    Constituent: 1% Fetal calf serum
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    80

  • Isotype

    IgG1

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8068 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 1/20.

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

 
IHC-Fr Use at an assay dependent concentration. Stains all types of keratin containing cells (epithelia) in frozen sections of various tissues, with the exception of myoepithelial cells.
IHC-P Use at an assay dependent concentration. For paraffin embedded tissue a TUF pretreatment is recommended.
WB Use at an assay dependent concentration. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Relevance

    Cytokeratin is a family of basic and acidic proteins, present in dermal tissues. Each cytokeratin is formed by heterotetramers of different types of keratin, that span from 1 to 18. In keratinized epidermis, 50 kD keratin is present in the basal layer, while 56.5 kD keratin is present in suprabasal layers, where as 58 kD keratin is present in the basal and suprabasal layers, while 65 to 67 kD keratin is present in the cells above the basal layers.

  • Cellular localization

    Cytoplasmic

  • Database links

    • Alternative names

      • CK1 antibody
      • Cytokeratin 1 antibody
      • EHK1 antibody
      • K1 antibody
      • Keratin 1 antibody
      • KRT1 antibody
      • KRT1A antibody
      • KRTA antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Keratin antibody [80] (ab8068)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Keratin antibody [80] (ab8068)This image is courtesy of an anonymous Abreview

      ab8068 staining human foreskin tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin embedded). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval in 10mM Citrate buffer pH 6.0 in boiling water bath for 10 minutes, followed by cooling at room temperature for 30 minutes. The blocking was done using 1%BSA/5% normal donkey serum in PBS pH 7.4 for 2 hours at room temperature. The primary antibody, diluted 1/10 (PBS pH 7.4, 1%BSA, 0.1% sodium azide) and incubated with sample for 16 hours at 4°C. A HRP-conjugated donkey polyclonal to mouse Ig, diluted 1/1000 was used as the secondary.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Keratin antibody [80] (ab8068)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Keratin antibody [80] (ab8068)
      ICC/IF image of ab8068 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8068, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Keratin antibody [80] (ab8068)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Keratin antibody [80] (ab8068)This image is courtesy of an anonymous Abreview

      Ab8068 staining pan Keratin in Human glioblastoma cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). The cells were fixed with paraformaldehyde; permeabilized with 0.1% Triton X 100 in PBS and blocked with 0.5% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody (1/50 dilution in 0.5% BSA in PBS) for 16hours at 4°C. An Alexa Fluor® 488 anti-mouse was used as a secondary antibody at 1/400 dilution.

      See Abreview

    • Western blot - Anti-pan Keratin antibody [80] (ab8068)
      Western blot - Anti-pan Keratin antibody [80] (ab8068)
      Anti-pan Keratin antibody [80] (ab8068) at 1/250 dilution + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

      Secondary
      Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 65 kDa
      Observed band size: 65 kDa
      Additional bands at: 49 kDa, 58 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 30 seconds
    • Flow Cytometry - Anti-pan Keratin antibody [80] (ab8068)
      Flow Cytometry - Anti-pan Keratin antibody [80] (ab8068)
      Overlay histogram showing A431 cells stained with ab8068 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 followed by the antibody (ab8068, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Keratin antibody [80] (ab8068)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Keratin antibody [80] (ab8068)Image courtesy of an anonymous Abreview.
      ab8068 staining pan Keratin in rat glioblastoma cell line C6 by Immunocytochemistry/ Immunofluorescence.
      Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab8068 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a Cy3 conjugated goat anti-mouse polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.

      See Abreview

    References

    This product has been referenced in:

    See all 22 Publications for this product

    Publishing research using ab8068? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 18 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-pan Keratin antibody [80]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Hair follicle)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH=6
    Permeabilization
    No
    Specification
    Hair follicle
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 07 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-pan Keratin antibody [80]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Keratinocytes (HaCaT))
    Permeabilization
    No
    Specification
    Keratinocytes (HaCaT)
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 07 2020

    Immunocytochemistry abreview for Anti-pan Keratin antibody [80]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cell (Keratinocytes (HaCaT))
    Permeabilization
    No
    Specification
    Keratinocytes (HaCaT)
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: PBS°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 07 2020

    Question

    I was wondering whether the abcam 8068 pan Keratin antibody might also work for rabbit tissue. More specifically we would like to use this antibody to stain the cornea which is mainly positive for Keratin 3 and 12 (also for CK 14 and 15). Would this Ab be able to detect (Cyto)Keratin 3 and 12?

    Read More
    Answer

    We are not aware of any tests of rabbit samples with the pan-keratin antibody ab8068. The production and initial characterization of the antibody are described in the two 1984 papers at these links.


    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1900394/

    https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1900469/


    I think the designation of the antibody as "pan-keratin" is based on the following comment from the Discussion section of the Jan 1984 paper, but we have not seen data from tests of purified keratins 3, 12, or others, or data from studies of cornea, so we can't say for sure that the antibody will be capable of detecting those keratins.


    It is clear from our staining results on normal tissues and malignant epithelial tumors that Clone 80 is a very broadly reacting antibody directed against an antigenic determinant common to a number of cytokeratins present in almost all epithelial cells.

    Read More

    Immunohistochemistry (Frozen sections) abreview for Anti-pan Keratin antibody [80]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Skin (psoriasis))
    Permeabilization
    No
    Specification
    Skin (psoriasis)
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 21°C
    Fixative
    Acetone
    Read More

    Dr. Christian Ostalecki

    Verified customer

    Submitted May 24 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-pan Keratin antibody [80]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (human glioblastoma cell line)
    Specification
    human glioblastoma cell line
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0,1% Triton X 100 in PBS
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 26 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-pan Keratin antibody [80]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (glioblastoma cell line C6)
    Specification
    glioblastoma cell line C6
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0,1% Triton X 100 in PBS
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 26 2012

    Question

    I am looking to get some prepared control slides for the Anti-pan Keratin antibody [80] (ab8068) on your website. The overview of your antibody product suggests that the positive control is Human Callus or keratinocytes. I am looking to purchase prepared slides of the suggested control and was curious if you offered such a product or could point me in the right direction? Additionally, how much would these slides possibly cost?   Thank you for your time and assistance.    

    Read More
    Answer

    Thank you for contacting Abcam. I have looked through our catalogue to see if we have any prepared slides of human callus or keratinocytes. I have not been able to find anything that matches your requirements, but I have placed a link below that will show all of the prepared slides that we do have: www.abcam.com/index.html?pageconfig=catalog_byclass&intClassID=4186&strStartChar=A,B I also did a quick search of other companies to see if they had anything that matches your requirements but I could not find anything. Please let me know if there is anything else I can help with.  

    Read More

    Question

    I came across primary antibody ab51054. Has this antibody ever been tested for its cross-reactivity against pig??? Thank you so much for all your help. Sincerely,

    Read More
    Answer

    Thank you for your question. We do not have any information about cross reactivity with pig. Please do not hesitate to contact us in case of any additional questions or remarks.

    Read More

    Question

    Thank you so much for the quick reply. I have another question. For my particular experiment I am co-culturing porcine periodontal ligament epithelial cells with mouse fibroblast cells. I need a primary antibody to distinguish these two different cells from one another. (mouse fibroblast cells have green fluorescent protein so double immunostaining is not necessary). Could you please recommend me with a primary antibody (preferably host animal to be a rabbit please) for this co-culture experiment. (my main goal is to distinguish two cells from each other). Thank you so much for all your help. Sincerely,

    Read More
    Answer

    Thank you for getting back to me and for explaining a little bit more about your experiments. It may be worth applying antibodies which recognize pig albumin or serum (ab79960, ab50337, ab79957). Unfortunately, currently we do not have many antibodies specific for porcine proteins. The majority of our antibodies has been routinely tested in human, mouse and rat. You could use antibodies against mouse MHC (class I) to detect and distinguish mouse cells from porcine cells. We have different clonality antibodies against mouse MHC class I, please have a look and use our new search engine. You can also filter through the search results by different criteria (such as application, reactivity, clonality): https://www.abcam.com/ I hope this helps and if I can assist further, please do not hesitate to contact me.

    Read More

    1-10 of 18 Abreviews or Q&A

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