重组Anti-Cytokeratin 8抗体[EP1628Y] - Cytoskeleton Marker (ab53280)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1628Y] to Cytokeratin 8 - Cytoskeleton Marker
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF, IHC-Fr
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Cytokeratin 8抗体[EP1628Y] - Cytoskeleton Marker
参阅全部 Cytokeratin 8 一抗 -
描述
兔单克隆抗体[EP1628Y] to Cytokeratin 8 - Cytoskeleton Marker -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF, IHC-Frmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IHC-P: Human breast adenocarcinona, ovarian carcinoma, breast carcinoima, colon adenocarcinoma, endometrial carcinoma and thyroid carcinoma tissue; mouse liver tissue; ICC/IF: HT-29 and HeLa cells; WB: HeLa, A431 and HaCaT cell lysates; Human breast cancer lysates and Mouse colon lysate; Flow Cyt (intra): HeLa cells, NIH/3T3 cells.
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常规说明
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
解离常数(KD)
KD = 4.60 x 10 -10 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1628Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-Cytokeratin 8 antibody [EP1628Y] (ab192467)
- Alexa Fluor® 647 Anti-Cytokeratin 8 antibody [EP1628Y] (ab192468)
- HRP Anti-Cytokeratin 8 antibody [EP1628Y] (ab193094)
- PE Anti-Cytokeratin 8 antibody [EP1628Y] (ab209297)
- Alexa Fluor® 405 Anti-Cytokeratin 8 antibody [EP1628Y] (ab210139)
- Anti-Cytokeratin 8 antibody [EP1628Y] - BSA and Azide free (ab217173)
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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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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab53280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。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 54 kDa).
For unpurified use at 1/25,000 - 1/50,000. |
IP |
1/20.
For unpurified use at 1:70. |
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IHC-P | (3) |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/100 - 1/500.
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IHC-Fr | (1) |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 54 kDa). For unpurified use at 1/25,000 - 1/50,000. |
IP
1/20. For unpurified use at 1:70. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/500. |
IHC-Fr
Use at an assay dependent concentration. |
靶标
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功能
Together with KRT19, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle. -
组织特异性
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. -
疾病相关
Cirrhosis -
序列相似性
Belongs to the intermediate filament family. -
翻译后修饰
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. -
细胞定位
Cytoplasm. Nucleus, nucleoplasm. Nucleus matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 3856 Human
- Entrez Gene: 16691 Mouse
- Entrez Gene: 25626 Rat
- Omim: 148060 Human
- SwissProt: P05787 Human
- SwissProt: P11679 Mouse
- SwissProt: Q10758 Rat
- Unigene: 533782 Human
see all -
别名
- CARD2 antibody
- CK 8 antibody
- CK-8 antibody
see all
图片
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Flow Cytometry (Intracellular) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Flow cytometry overlay histogram showing left NIH3T3 positive cells and right negative Raw264.7 stained with ab53280 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab53280) (1x 106 in 100μl at 0.2μg/ml (1/11500)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) at 1/10000 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.
Predicted band size: 54 kDaLanes 1 - 4: Merged signal (red and green). Green - ab53280 observed at 55 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab53280 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. ab53280 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 (For unpurified use at 1/25,000 - 1/50,000) dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Immunocytochemistry/ Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling Cytokeratin 9 with Purified ab53280 at 1:500 dilution. Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Alexa Fluor® 488 (ab192467) and Alexa Fluor® 647 (ab192468) conjugated versions are available for this clone.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling Cytokeratin 8 with Purified ab53280 at 1:250 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Flow Cytometry (Intracellular) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 8 with purified ab53280 at 1/20 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Alexa Fluorr®488 (ab192467) and Alexa Fluorr®647 (ab192468) conjugated versions are available for this clone.
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All lanes : Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) at 1/10000 dilution (purified)
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates
Lane 2 : Human breast cancer lysates
Lane 3 : HaCaT (Human skin keratinocyte) whole cell lysates
Lane 4 : Mouse colon lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 54 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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ab53280 (purified) at 1:20 dilution (0.2μg) immunoprecipitating Cytokeratin 8 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10μg
Lane 2 (+): ab53280 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab53280 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)Stewart, M.K. et al PLoS One. 2016 Apr 21;11(4):e0154162. doi: 10.1371/journal.pone.0154162. eCollection 2016 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Panx1-/- mice have normal mammary gland epithelial differentiation at lactation
Immunofluorescent analysis of luminal epithelial marker keratin 8 (green) and myoepithelial marker keratin14 (red) revealed a similar staining pattern in Panx1-/- mice compared to control mice during lactation. Paraffin-embedded tissue samples.
Hoescht (blue) denotes nuclei. N = 6. Scale bars = 50 um.
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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Cytokeratin 8 with purified ab53280 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Unpurified ab53280 (1:250) staining human Cyotkeratin 8 in human breast adenocarcinoma tissue by immunohistochemistry using paraffin embedded tissue.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Cytokeratin 8 with Purified ab53280 at 1:250 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280) at 1/50000 dilution (unpurified) + A431 cell lysate at 10 µg
Secondary
Goat anti-Rabbit HRP labeled at 1/2000 dilution
Predicted band size: 54 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)This image is courtesy of an anonymous Abreview
Unpurified ab53280 staining Cytokeratin 8 in Mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formalin and blocked with 10% serum for 20 minutes at 23°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/75 in TBS + 1% BSA) for 1 hour at 23°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Immunofluorescent staining of HeLa cells using unpurified ab53280 (1:100).
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Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)This image is courtesy of Dr. Shaohua Li
Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School
Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)
Preparation:
Fix in 3% PFA in PBS for 30 min at RT Incubate in 7.5% sucrose-PBS for 3h at RT Incubate in 15% sucrose-PBS at 4 degree Celsius overnight Embed the EBs in tissue-Tek OCT compound Cut frozen sections to 4-20 µm thickness
Primary antibody 1: Rabbit anti cytokeratin 8 (unpurified ab53280), 1:100
Primary antibody 2: Rat anti-perlecan, 1:100
Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed (ab150081), 1:200
Nuclei were counterstained with DAPI -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Fluorescent immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue using unpurified ab53280. Green-CK8 red-PI.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab53280. Green-CK8 red-PI
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Fluorescent immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue using unpurified ab53280. Green-CK8 red-PI.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Fluorescent immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue using unpurified ab53280. Green-CK8 red-PI.
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Flow Cytometry (Intracellular) - Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker (ab53280)
Overlay histogram showingHeLa cells stained with unpurifiedab53280 (red line). The cells were fixed with 2%PFA (room temperature, 30 min) and then permeabilized with 1%FACS permeabilizing solutionfor 30 min. The cells were then incubated in3% FBS in 1X PBSfollowed by the antibody (ab53280, 1/20 dilution) for1 hourat room temperature. The cells were then incubated for 30 min at room temperature with the secondary antibody. An isotype control antibody (black line) was used and an unlabelled sample (blue line) was also used as a control.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (105)
ab53280 被引用在 105 文献中.
- Wang W et al. Targeted Deletion of Kindlin-2 in Mouse Mammary Glands Inhibits Tumor Growth, Invasion, and Metastasis Downstream of a TGF-β/EGF Oncogenic Signaling Pathway. Cancers (Basel) 14:N/A (2022). PubMed: 35158908
- Marhold M et al. MALAT1 Fusions and Basal Cells Contribute to Primary Resistance against Androgen Receptor Inhibition in TRAMP Mice. Cancers (Basel) 14:N/A (2022). PubMed: 35159020
- Sheftel CM et al. Peripartal treatment with low-dose sertraline accelerates mammary gland involution and has minimal effects on maternal and offspring bone. Physiol Rep 10:e15204 (2022). PubMed: 35234346
- Yang L et al. Comprehensive Analysis of the Transcriptome-Wide m6A Methylome in Endometrioid Ovarian Cancer. Front Oncol 12:844613 (2022). PubMed: 35280730
- Gopinathan G et al. Immune Mechanisms of Resistance to Cediranib in Ovarian Cancer. Mol Cancer Ther 21:1030-1043 (2022). PubMed: 35313341