Recombinant Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab68459)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1619Y] to Cytokeratin 7 - Cytoskeleton Marker
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker
See all Cytokeratin 7 primary antibodies -
Description
Rabbit monoclonal [EPR1619Y] to Cytokeratin 7 - Cytoskeleton Marker -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Cytokeratin 7 aa 1-100 (N terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa, SK-OV-3 and T47D cell lysates. IHC-P: Human ovarian carcinoma and human bladder carcinoma tissues. ICC/IF: HeLa and TD47 cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate (ab150035).
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 2.10 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1619Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-Cytokeratin 7 antibody [EPR1619Y] - BSA and Azide free (ab181831)
- Alexa Fluor® 488 Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab185048)
- Alexa Fluor® 647 Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab192077)
- HRP Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab192079)
- Alexa Fluor® 594 Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab203427)
- Alexa Fluor® 555 Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab203434)
- PE Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab210628)
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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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68459 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 |
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Flow Cyt (Intra) |
1/20 - 1/30.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/5000 - 1/10000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
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IP |
1/20 - 1/30.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
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ICC/IF | (1) |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/20 - 1/30. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/5000 - 1/10000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa). |
IP
1/20 - 1/30. |
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
ICC/IF
1/100 - 1/250. |
Target
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Function
Blocks interferon-dependent interphase and stimulates DNA synthesis in cells. Involved in the translational regulation of the human papillomavirus type 16 E7 mRNA (HPV16 E7). -
Tissue specificity
Expressed in cultured epidermal, bronchial and mesothelial cells but absent in colon, ectocervix and liver. Observed throughout the glandular cells in the junction between stomach and esophagus but is absent in the esophagus. -
Sequence similarities
Belongs to the intermediate filament family. -
Post-translational
modificationsArg-20 is dimethylated, probably to asymmetric dimethylarginine. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 3855 Human
- Omim: 148059 Human
- SwissProt: P08729 Human
- Unigene: 411501 Human
- Unigene: 670221 Human
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Alternative names
- CK 7 antibody
- CK-7 antibody
- CK7 antibody
see all
Images
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All lanes : Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab68459) at 1/5000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : KRT7 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 51 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab68459 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab68459 was shown to specifically react with KRT7 (Cytokeratin 7) in wild-type A549 cells as signal was lost in KRT7 knockout cells. Wild-type and KRT7 knockout samples were subjected to SDS-PAGE. Ab68459 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Cytokeratin 7 with unpurified ab68459.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of T47D cells labelling Cytokeratin 7 with purified ab68459 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
Alexa Fluor® 488 (ab185048) and Alexa Fluor® 647 (ab192077) conjugated versions are available for this clone.
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All lanes : Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab68459) at 1/10000 dilution (purified)
Lane 1 : HeLa whole cell lysate
Lane 2 : SK-OV-3 whole cell lysate
Lane 3 : T47D whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 51 kDa
Observed band size: 51 kDa
Blocking and dilution buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling Cytokeratin 7 with purified ab68459 at a dilution of 1/1000. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Intracellular Flow Cytometry analysis of HeLa cells labelling Cytokeratin 7 with purified ab68459 at a dilution of 1/20 (red). Cells were fixed with 80% methanol. An Alexa Fluorr®488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Alexa Fluorr®488 (ab185048) and Alexa Fluorr®647 (ab192077) conjugated versions are available for this clone.
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ab68459 (purified) at 1/20 immunoprecipitating Cytokeratin 7 in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab68459 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68459 in HeLa whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab68459) at 1/10000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : T47D cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 51 kDa
Observed band size: 51 kDa -
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Cytokerain 7 with unpurified ab68459 at a dilution of 1/100.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrial carcinoma tissue labelling Cytokeratin 7 with unpurified ab68459.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling Cytokeratin 7 with unpurified ab68459.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human urinary bladder transitional carcinoma tissue labelling Cytokeratin 7 with unpurified ab68459.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab68459 showing negative staining in human sarcoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab68459 showing negative staining in human colonic adenocarcinoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing HeLa cells stained with unpurified ab68459 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% 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 (unpurified ab68459, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (31)
ab68459 has been referenced in 31 publications.
- Li G et al. Endometrial stromal cell ferroptosis promotes angiogenesis in endometriosis. Cell Death Discov 8:29 (2022). PubMed: 35039492
- Zhang W et al. Rif1 and Hmgn3 regulate the conversion of murine trophoblast stem cells. Cell Rep 38:110570 (2022). PubMed: 35354046
- Bai Z et al. Lapatinib Suppresses HER2-Overexpressed Cholangiocarcinoma and Overcomes ABCB1- Mediated Gemcitabine Chemoresistance. Front Oncol 12:860339 (2022). PubMed: 35463361
- Murrow LM et al. Mapping hormone-regulated cell-cell interaction networks in the human breast at single-cell resolution. Cell Syst 13:644-664.e8 (2022). PubMed: 35863345
- Osnato A et al. TGFß signalling is required to maintain pluripotency of human naïve pluripotent stem cells. Elife 10:N/A (2021). PubMed: 34463252