製品の概要

  • 製品名
    Anti-pan Keratin antibody [80]
    pan Keratin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [80] to pan Keratin
  • アプリケーション
    適用あり: Flow Cyt, IHC-Fr, IHC-P, WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Callus cytokeratins isolated from fresh human skin tissue

  • ポジティブ・コントロール
    • Human callus or keratinocytes

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    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.
  • バッファー
    Preservative: 0.07% Sodium azide
    Constituent: 1% Fetal calf serum
  • 精製度
    Tissue culture supernatant
  • ポリ/モノ
    モノクローナル
  • クローン名
    80
  • アイソタイプ
    IgG1
  • 研究分野

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 関連性
    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.
  • 細胞内局在
    Cytoplasmic
  • 参照データベース
  • 別名
    • CK1 antibody
    • Cytokeratin 1 antibody
    • EHK1 antibody
    • K1 antibody
    • Keratin 1 antibody
    • KRT1 antibody
    • KRT1A antibody
    • KRTA antibody
    see all

画像

  • 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

  • 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.

  • 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

  • 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
  • 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.

  • 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

参考文献

This product has been referenced in:
  • Jiang Z  et al. LincIN, a novel NF90-binding long non-coding RNA, is overexpressed in advanced breast tumors and involved in metastasis. Breast Cancer Res 19:62 (2017). Read more (PubMed: 28558830) »
  • Cao L  et al. Keratin mediates the recognition of apoptotic and necrotic cells through dendritic cell receptor DEC205/CD205. Proc Natl Acad Sci U S A 113:13438-13443 (2016). Read more (PubMed: 27821726) »

See all 16 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 Jan 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 Jan 26 2012

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 s...

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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.

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 anti...

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Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this product. As our Abpromise indicates, in the event that a prod...

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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 (foreskin)
Specification
foreskin
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate pH 6
Permeabilization
No
Blocking step
1%BSA plus 5% normal donkey serum in PBS pH 7.4 as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Sep 30 2008

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (epithelial)
Specification
epithelial
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1%
Username

Dr. todd french

Verified customer

投稿 Feb 20 2007

Thank you for your enquiry. When conducting IHC, it is not advisable to use antibodies which were raised in the same species as the target sample e.g. a mouse mAb against mouse sample. The secondary Ab may recognise endogenous IgG in your sample tis...

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Unfortunately, we are not specialists in this area and we do not want to recommend a product that is unsuitable for your needs. We suggest that you refer to the latest literature in this field.

1-10 of 13 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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