Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker

• 产品名称

  Anti-Cytokeratin 8抗体[M20] - Cytoskeleton Marker
  参阅全部 Cytokeratin 8 一抗

• 描述

  小鼠单克隆抗体[M20] to Cytokeratin 8 - Cytoskeleton Marker

• 宿主

  Mouse

• 经测试应用

  适用于: Flow Cyt, ICC, ICC/IF, IHC-P, IHC-Fr, WBmore details

• 种属反应性

  与反应: Rat, Rabbit, Human
  

• 免疫原

  Full length native protein (purified) corresponding to Human Cytokeratin 8. Keratin isolated from the human breast carcinoma cell line MCF-7.

• 阳性对照

• WB: HeLa, A431, PC3 and LNCaP cell lysates. ICC/IF: MCF-7, pancreatic cancer and epithelial ovarian cancer cells. IHC-Fr: Human colon tissue.

• 常规说明

  Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 – 7.8. The individual cytokeratin polypeptides are numbered 1 to 20. The various epithelia in the human body usually express cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. The cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C.

• 存储溶液

  pH: 7.3
  Preservative: 0.1% Sodium azide
  Constituents: PBS, 1% Fetal calf serum

• 纯度

  Tissue culture supernatant

• Primary antibody说明

  Cytokeratins are a subfamily of intermediate filament proteins and are characterized by a remarkable biochemical diversity, represented in epithelial tissues by at least 20 different polypeptides. They range in molecular weight between 40 kDa and 68 kDa and isoelectric pH between 4.9 – 7.8. The individual cytokeratin polypeptides are numbered 1 to 20. The various epithelia in the human body usually express cytokeratins which are not only characteristic of the type of epithelium, but also related to the degree of maturation or differentiation within an epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. The cytokeratin antibodies are not only of assistance in the differential diagnosis of tumors using immunohistochemistry on tissue sections, but are also a useful tool in cytopathology and flow cytometric assays.

• 克隆

  单克隆

• 克隆编号

  M20

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab9023于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

数据库链接

• Entrez Gene: 3856 Human

• Entrez Gene: 100348891 Rabbit

• Entrez Gene: 25626 Rat

• Omim: 148060 Human

• SwissProt: P05787 Human

• SwissProt: Q10758 Rat

• Unigene: 533782 Human

• Unigene: 708445 Human

• Unigene: 11083 Rat

别名

• CARD2 antibody

• CK 8 antibody

• CK-8 antibody

• CK8 antibody

• CYK8 antibody

• CYKER antibody

• Cytokeratin endo A antibody

• Cytokeratin-8 antibody

• DreK8 antibody

• EndoA antibody

• K2C8 antibody

• K2C8_HUMAN antibody

• K8 antibody

• Keratin 8 antibody

• Keratin type II cytoskeletal 8 antibody

• Keratin, type II cytoskeletal 8 antibody

• Keratin-8 antibody

• KO antibody

• Krt 2.8 antibody

• KRT8 antibody

• MGC118110 antibody

• MGC174782 antibody

• MGC53564 antibody

• MGC85764 antibody

• sb:cb186 antibody

• Type-II keratin Kb8 antibody

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  Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  All lanes : Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023) at 1/1000 dilution
  
  Lane 1 : A431 cell lysate
  Lane 2 : MCF7 cell lysate
  Lane 3 : Wild-type HeLa cell lysate
  Lane 4 : KRT8 knockout HeLa cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  
  

  Lanes 1 - 4: Merged signal (red and green). Green - ab9023 observed at 55 kDa. Red - loading control, ab181602 observed at 37 kDa.  

   ab9023 was shown to react with Cytokeratin 8 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255400 (knockout cell lysate ab263785) was used. Wild-type and Cytokeratin 8 knockout samples were subjected to SDS-PAGE. ab9023 and Anti-GAPDH antibody EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

   

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  Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  The image shows different concentration of total protein from normal and tumour colon tissues.

  This picture was kindly supplied as part of the review submitted by Semona Rupchand.

  The image shows different concentration of total protein from normal and tumour colon tissues.

  This picture was kindly supplied as part of the review submitted by Semona Rupchand.

• 

  Western blot - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)This image is a courtesy of Anonymous Abreview

  All lanes : Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023) at 1/2000 dilution
  
  Lane 1 : Lysate prepared from human prostate cancer LNCaP cells
  Lane 2 : Lysate prepared from human prostate cancer PC3 cells
  
  Lysates/proteins at 50 µg per lane.
  
  Secondary
  All lanes : HRP-conjugated goat polyclonal to mouse IgG at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Observed band size: 50 kDawhy is the actual band size different from the predicted?
  Additional bands at: 26 kDa, 65 kDa. We are unsure as to the identity of these extra bands.
  
  
  Exposure time: 4 minutes
  

  See Abreview

• 

  Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  Immunohistochemistry analysis of frozen section of human colon labeling Cytokeratin 8 with ab9023.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  Immunohistochemistry on paraffin section of human colon

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  Ab9023 staining human normal liver parenchima tissue. Staining is localized to cell membrane.
  Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
  Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

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  Flow Cytometry - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  Overlay histogram showing MCF-7 cells stained with ab9023 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.5% 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 followed by the antibody (ab9023, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/250 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. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.5% PBS-Tween used under the same conditions.

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  Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker (ab9023)

  Immunohistochemistry on frozen section of human colon