Recombinant Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)](https://www.abcam.com/ps/products/51/ab51599/Images/ab51599-63525-anti-cytokeratin-4-antibody-ep1599y-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1599Y] to Cytokeratin 4
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cytokeratin 4 antibody [EP1599Y]
See all Cytokeratin 4 primary antibodies -
Description
Rabbit monoclonal [EP1599Y] to Cytokeratin 4 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Cytokeratin 4 (C terminal). The exact sequence is proprietary.
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Positive control
- A431 cell lysate and human cervical carcinoma tissue. This antibody gave a positive result when used in the following methanol fixed cell lines: A431 Flow Cyt (intra): HeLa cells IP: HaCaT whole cell lysate.
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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 -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1599Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab51599 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/100 - 1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| ICC/IF |
1/50 - 1/100.
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| IP |
1/10 - 1/100.
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| WB |
1/1000 - 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 57 kDa).
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| IHC-P | (1) |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Notes |
|---|
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Flow Cyt (Intra)
1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF
1/50 - 1/100. |
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IP
1/10 - 1/100. |
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WB
1/1000 - 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 57 kDa). |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Tissue specificity
Detected in the suprabasal layer of the stratified epithelium of the esophagus, exocervix, vagina, mouth and lingual mucosa, and in cells and cell clusters in the mucosa and serous gland ducts of the esophageal submucosa (at protein level). Expressed widely in the exocervix and esophageal epithelium, with lowest levels detected in the basal cell layer. -
Involvement in disease
Defects in KRT4 are a cause of white sponge nevus of cannon (WSN) [MIM:193900]. WSN is a rare autosomal dominant disorder which predominantly affects non-cornified stratified squamous epithelia. Clinically, it is characterized by the presence of soft, white, and spongy plaques in the oral mucosa. The characteristic histopathologic features are epithelial thickening, parakeratosis, and vacuolization of the suprabasal layer of oral epithelial keratinocytes. Less frequently the mucous membranes of the nose, esophagus, genitalia and rectum are involved. -
Sequence similarities
Belongs to the intermediate filament family. - Information by UniProt
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Database links
- Entrez Gene: 3851 Human
- Omim: 123940 Human
- SwissProt: P19013 Human
- Unigene: 654610 Human
- Unigene: 706527 Human
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Form
Localisation: Intermediate filament (Cytoskeleton). -
Alternative names
- CK 4 antibody
- CK-4 antibody
- CK4 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)
Immunohistochemical staining of paraffin embedded human cervical carcinoma tissue using ab51599 (1/100). Goat anti-rabbit HRP (1/2000) used for secondary staining.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)](https://www.abcam.com/ps/products/51/ab51599/Images/ab51599-219237-anti-cytokeratin-4-antibody-ep1599y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)ICC/IF image of ab51599 stained A431 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab51599 at 1/100 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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![Western blot - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)](https://www.abcam.com/ps/products/51/ab51599/Images/ab51599-63526-anti-cytokeratin-4-antibody-ep1599y-western-blot.jpg)
Western blot - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599) at 1/1000 dilution + A431 cell lysate at 10 µg
Secondary
Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 57 kDa
Observed band size: 59 kDa why is the actual band size different from the predicted? -
![Immunoprecipitation - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)](https://www.abcam.com/ps/products/51/ab51599/Images/ab51599-6-anti-cytokeratin-4-antibody-ep1599y-immunoprecipitation-hacat-human.jpg)
Immunoprecipitation - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)Purified ab51599 at 1/50 dilution (2µg) immunoprecipitating Cytokeratin 4 in HaCaT whole cell lysate.
Lane 1 (input): HaCaT (Human skin keratinocyte) whole cell lysate 10µg
Lane 2 (+): ab51599 + HaCaT whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51599 in HaCaT whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 59 kDa -
![Flow Cytometry (Intracellular) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)](https://www.abcam.com/ps/products/51/ab51599/Images/ab51599-5-anti-cytokeratin-4-antibody-ep1599y-flowcytometry-hela-human.jpg)
Flow Cytometry (Intracellular) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 4 with purified ab51599 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).
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![Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)](https://www.abcam.com/ps/products/51/ab51599/Images/ab51599-4-benefits-of-recombinant-antibodies.png)
Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)
Protocols
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.
Datasheets and documents
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Datasheet download
References (12)
ab51599 has been referenced in 12 publications.
- Luo Q et al. TRIM32/USP11 Balances ARID1A Stability and the Oncogenic/Tumor-Suppressive Status of Squamous Cell Carcinoma. Cell Rep 30:98-111.e5 (2020). PubMed: 31914402
- Hu L et al. Human esophageal myofibroblasts increase squamous epithelial thickness via paracrine mechanisms in an in vitro model of gastroesophageal reflux disease. PLoS One 15:e0238852 (2020). PubMed: 32925965
- Luo Q et al. ARID1A prevents squamous cell carcinoma initiation and chemoresistance by antagonizing pRb/E2F1/c-Myc-mediated cancer stemness. Cell Death Differ 27:1981-1997 (2020). PubMed: 31831874
- Yokoo S & Yamagami S Goblet Cell Differentiation Potential in Human Corneal Limbal Epithelial Progenitor Cells In Vitro. Invest Ophthalmol Vis Sci 61:27 (2020). PubMed: 33112944
- Zhang C et al. Human esophageal myofibroblast secretion of bone morphogenetic proteins and GREMLIN1 and paracrine regulation of squamous epithelial growth. Sci Rep 8:12354 (2018). PubMed: 30120338
