Anti-Cytokeratin 8 抗体 [M20] - Cytoskeleton Marker (ab9023)
Key features and details
- Mouse monoclonal [M20] to Cytokeratin 8 - Cytoskeleton Marker
- Suitable for: Flow Cyt, ICC, ICC/IF, IHC-P, IHC-Fr, WB
- Knockout validated
- Reacts with: Rat, Rabbit, Human
- Isotype: IgG1
製品の概要
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製品名
Anti-Cytokeratin 8 antibody [M20] - Cytoskeleton Marker
Cytokeratin 8 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [M20] to Cytokeratin 8 - Cytoskeleton Marker -
由来種
Mouse -
アプリケーション
適用あり: Flow Cyt, ICC, ICC/IF, IHC-P, IHC-Fr, WBmore details -
種交差性
交差種: 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.
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ポジティブ・コントロール
- WB: HeLa, A431, PC3 and LNCaP cell lysates. ICC/IF: MCF-7, pancreatic cancer and epithelial ovarian cancer cells. IHC-Fr: Human colon tissue.
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特記事項
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.
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製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. -
バッファー
pH: 7.3
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% Fetal calf serum -
Concentration information loading...
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精製度
Tissue culture supernatant -
一次抗体 備考
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. -
ポリ/モノ
モノクローナル -
クローン名
M20 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab9023の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt |
Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC | (1) |
Use at an assay dependent concentration.
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ICC/IF | (4) |
Use at an assay dependent concentration.
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IHC-P | (1) |
1/5 - 1/25.
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IHC-Fr |
1/100 - 1/200. For human colon 20min ice cold acetone fixation was carried out with 60min incubation with ab9023 at 37C.
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WB | (4) |
1/100 - 1/1000.
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特記事項 |
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Flow Cyt
Use 2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
1/5 - 1/25. |
IHC-Fr
1/100 - 1/200. For human colon 20min ice cold acetone fixation was carried out with 60min incubation with ab9023 at 37C. |
WB
1/100 - 1/1000. |
ターゲット情報
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機能
Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle. -
組織特異性
Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma membrane in structures that contain dystrophin and spectrin. Expressed in gingival mucosa and hard palate of the oral cavity. -
関連疾患
Cirrhosis -
配列類似性
Belongs to the intermediate filament family. -
翻訳後修飾
Phosphorylation on serine residues is enhanced during EGF stimulation and mitosis. Ser-74 phosphorylation plays an important role in keratin filament reorganization.
O-glycosylated. O-GlcNAcylation at multiple sites increases solubility, and decreases stability by inducing proteasomal degradation.
O-glycosylated (O-GlcNAcylated), in a cell cycle-dependent manner. -
細胞内局在
Cytoplasm. Nucleus, nucleoplasm. Nucleus matrix. - Information by UniProt
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参照データベース
- 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
see all -
別名
- CARD2 antibody
- CK 8 antibody
- CK-8 antibody
see all
画像
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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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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.
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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 kDa why 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
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Immunohistochemistry analysis of frozen section of human colon labeling Cytokeratin 8 with ab9023.
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Immunohistochemistry on paraffin section of human colon
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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. -
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 on frozen section of human colon
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (34)
ab9023 は 34 報の論文で使用されています。
- Ryosaka M et al. Protocol for the generation and expansion of human iPS cell-derived ureteric bud organoids. STAR Protoc 3:101484 (2022). PubMed: 35769929
- Baek A et al. KRT8 (keratin 8) attenuates necrotic cell death by facilitating mitochondrial fission-mediated mitophagy through interaction with PLEC (plectin). Autophagy 17:3939-3956 (2021). PubMed: 33783309
- Lee SY et al. Reducing tumor invasiveness by ramucirumab and TGF-β receptor kinase inhibitor in a diffuse-type gastric cancer patient-derived cell model. Cancer Med 10:7253-7262 (2021). PubMed: 34542244
- Liu Z et al. Wnt ligands 3a and 5a regulate proliferation and migration in human fetal liver progenitor cells. Transl Gastroenterol Hepatol 6:56 (2021). PubMed: 34805578
- Scott MKD et al. A multi-scale integrated analysis identifies KRT8 as a pan-cancer early biomarker. Pac Symp Biocomput 26:297-308 (2021). PubMed: 33691026