Recombinant Anti-Cytokeratin 19 antibody [EPR1579Y] (ab76539)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1579Y] to Cytokeratin 19
- 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 19 antibody [EPR1579Y]
See all Cytokeratin 19 primary antibodies -
Description
Rabbit monoclonal [EPR1579Y] to Cytokeratin 19 -
Host species
Rabbit -
Specificity
In our WB testing this antibody did not recognize mouse Cytokeratin 19, but customers have reported the antibody recognizes the protein in IHC-P with mouse tissue. Thus, we have removed mouse as a "does not react with" species and welcome further feedback from other researchers using this antibody in mouse. -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: T47D, HepG2 or SKBR-3 cell lysate; IHC-P: Human stomach adenocarcinoma tissue. IF/ICC: HCT116 cell line. MCF7 cells (MCF7-KRT19 KO used as negative cell line).
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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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1579Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EPR1579Y] (ab205445)
- Alexa Fluor® 647 Anti-Cytokeratin 19 antibody [EPR1579Y] (ab205446)
- PE Anti-Cytokeratin 19 antibody [EPR1579Y] (ab216705)
- APC Anti-Cytokeratin 19 antibody [EPR1579Y] (ab221255)
- PerCP Anti-Cytokeratin 19 antibody [EPR1579Y] (ab221256)
- Anti-Cytokeratin 19 antibody [EPR1579Y] - BSA and Azide free (ab232566)
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76539 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.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/10000 - 1/50000. Predicted molecular weight: 44 kDa.
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IP |
1/20.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
|
ICC/IF | (1) |
1/100 - 1/250.
Signal can be observed in cells fixed with either methanol or paraformaldehyde. |
Notes |
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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 - 1/50000. Predicted molecular weight: 44 kDa. |
IP
1/20. |
IHC-P
1/1000. Perform heat mediated antigen retrieval via the pressure cooker method 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. Signal can be observed in cells fixed with either methanol or paraformaldehyde. |
Target
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Function
Involved in the organization of myofibers. Together with KRT8, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle. -
Tissue specificity
Expressed in a defined zone of basal keratinocytes in the deep outer root sheath of hair follicles. Also observed in sweat gland and mammary gland ductal and secretory cells, bile ducts, gastrointestinal tract, bladder urothelium, oral epithelia, esophagus, ectocervical epithelium (at protein level). Expressed in epidermal basal cells, in nipple epidermis and a defined region of the hair follicle. Also seen in a subset of vascular wall cells in both the veins and artery of human umbilical cord, and in umbilical cord vascular smooth muscle. Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma in structures that contain dystrophin and spectrin. -
Sequence similarities
Belongs to the intermediate filament family. -
Developmental stage
Present in hair follicles at all stages of development. -
Domain
This keratin differs from all other IF proteins in lacking the C-terminal tail domain. - Information by UniProt
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Database links
- Entrez Gene: 3880 Human
- Omim: 148020 Human
- SwissProt: P08727 Human
- Unigene: 654568 Human
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Alternative names
- 40 kDa keratin intermediate filament antibody
- CK 19 antibody
- CK-19 antibody
see all
Images
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ab76539 staining Cytokeratin 19 in wild-type MCF7 cells (top panel) and KRT19 knockout MCF7 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab76539 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling Cytokeratin 19 with Purified ab76539 at 1:1000 dilution (0.12 µg/ml). 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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ab76539 (purified) at 1:20 dilution (2ug) immunoprecipitating Cytokeratin 19 in SKBR-3 whole cell lysate.
Lane 1 (input): SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab76539 & SKBR-3 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76539 in SKBR-3 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. -
Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cytokeratin 19 with purified ab76539 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 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).
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All lanes : Anti-Cytokeratin 19 antibody [EPR1579Y] (ab76539) at 1/10000 dilution (unpurified)
Lane 1 : T47D cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : SKBR-3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 44 kDa -
Immunohistochemical staining of human stomach adenocarcinoma tissue with ab76539 (unpurified) at1/100-1/250.
Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
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ab76539 (unpurified) stained HCT116 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76539 at 1/200 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
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Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab76539 (unpurified). Green- CK19 red-PI.
Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
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Overlay histogram showing MCF7 cells stained with unpurified ab76539 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab76539, 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. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (11)
ab76539 has been referenced in 11 publications.
- Wang Z et al. Carcinomas assemble a filamentous CXCL12-keratin-19 coating that suppresses T cell-mediated immune attack. Proc Natl Acad Sci U S A 119:N/A (2022). PubMed: 35046049
- Capeloa T et al. MitoQ Prevents Human Breast Cancer Recurrence and Lung Metastasis in Mice. Cancers (Basel) 14:N/A (2022). PubMed: 35326639
- Del Valle JS et al. Dynamic in vitro culture of cryopreserved-thawed human ovarian cortical tissue using a microfluidics platform does not improve early folliculogenesis. Front Endocrinol (Lausanne) 13:936765 (2022). PubMed: 35966050
- Sheng W et al. Upregulation of Linc00284 Promotes Lung Cancer Progression by Regulating the miR-205-3p/c-Met Axis. Front Genet 12:694571 (2021). PubMed: 34616424
- Sun M et al. Cleavable cellulosic sponge for functional hepatic cell culture and retrieval. Biomaterials 201:16-32 (2019). PubMed: 30784769