Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1612Y] to Cytokeratin 14
- Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Product nameAnti-Cytokeratin 14 antibody [EP1612Y]
See all Cytokeratin 14 primary antibodies
DescriptionRabbit monoclonal [EP1612Y] to Cytokeratin 14
Tested applicationsSuitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Synthetic peptide within Human Cytokeratin 14 aa 400-500 (C terminal). The exact sequence is proprietary.
- WB: A431 cell lysate. IHC-P: Human skin tissue. ICC/IF: A431 cells. Flow Cyt: A431 cells.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
- Anti-Cytokeratin 14 antibody [EP1612Y] (Alexa Fluor® 488) (ab192055)
- Anti-Cytokeratin 14 antibody [EP1612Y] (Alexa Fluor® 647) (ab192056)
- Anti-Cytokeratin 14 antibody [EP1612Y] (HRP) (ab192081)
- Anti-Cytokeratin 14 antibody [EP1612Y] (PE) (ab210414)
- Anti-Cytokeratin 14 antibody [EP1612Y] (Alexa Fluor® 555) (ab214391)
- Anti-Cytokeratin 14 antibody [EP1612Y] - BSA and Azide free (ab243907)
Our Abpromise guarantee covers the use of ab51054 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/20000. Detects a band of approximately 48 kDa (predicted molecular weight: 52 kDa).|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionThe nonhelical tail domain is involved in promoting KRT5-KRT14 filaments to self-organize into large bundles and enhances the mechanical properties involved in resilience of keratin intermediate filaments in vitro.
Tissue specificityDetected in the basal layer, lowered within the more apically located layers specifically in the stratum spinosum, stratum granulosum but is not detected in stratum corneum. Strongly expressed in the outer root sheath of anagen follicles but not in the germinative matrix, inner root sheath or hair. Found in keratinocytes surrounding the club hair during telogen.
Involvement in diseaseDefects in KRT14 are a cause of epidermolysis bullosa simplex Dowling-Meara type (DM-EBS) [MIM:131760]. DM-EBS is a severe form of intraepidermal epidermolysis bullosa characterized by generalized herpetiform blistering, milia formation, dystrophic nails, and mucous membrane involvement.
Defects in KRT14 are a cause of epidermolysis bullosa simplex Weber-Cockayne type (WC-EBS) [MIM:131800]. WC-EBS is a form of intraepidermal epidermolysis bullosa characterized by blistering limited to palmar and plantar areas of the skin.
Defects in KRT14 are a cause of epidermolysis bullosa simplex Koebner type (K-EBS) [MIM:131900]. K-EBS is a form of intraepidermal epidermolysis bullosa characterized by generalized skin blistering. The phenotype is not fundamentally distinct from the Dowling-Meara type, although it is less severe.
Defects in KRT14 are the cause of epidermolysis bullosa simplex autosomal recessive (AREBS) [MIM:601001]. AREBS is an intraepidermal epidermolysis bullosa characterized by localized blistering on the dorsal, lateral and plantar surfaces of the feet.
Defects in KRT14 are the cause of Naegeli-Franceschetti-Jadassohn syndrome (NFJS) [MIM:161000]; also known as Naegeli syndrome. NFJS is a rare autosomal dominant form of ectodermal dysplasia. The cardinal features are absence of dermatoglyphics (fingerprints), reticular cutaneous hyperpigmentation (starting at about the age of 2 years without a preceding inflammatory stage), palmoplantar keratoderma, hypohidrosis with diminished sweat gland function and discomfort provoked by heat, nail dystrophy, and tooth enamel defects.
Defects in KRT14 are the cause of dermatopathia pigmentosa reticularis (DPR) [MIM:125595]. DPR is a rare ectodermal dysplasia characterized by lifelong persistent reticulate hyperpigmentation, noncicatricial alopecia, and nail dystrophy.
Sequence similaritiesBelongs to the intermediate filament family.
Cellular localizationCytoplasm. Nucleus. Expressed in both as a filamentous pattern.
- Information by UniProt
- CK 14 antibody
- CK-14 antibody
- ck14 antibody
All lanes : Anti-Cytokeratin 14 antibody [EP1612Y] (ab51054) at 1/10000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : KRT14 knockout A431 cell lysate
Lane 3 : Human skin cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green - ab51054 observed at 49 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab51054 was shown to react with Cytokeratin 14 in wild-type A431 cells in western blot. Loss of signal was observed when KRT14 knockout sample was used. Wild-type A431 and KRT14 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab51054 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
ab51054 staining Cytokeratin 14 in human skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer, pH 6.0. Samples were then permeabilized using 0.1% saponin/PBS, blocked with 4% BSA for 30 minutes at 25°C and then incubated with ab51054 at a 1/200 dilution for 16 hours at 4°C. The secondary used was a Texas Red conjugated goat anti-rabbit polyclonal used at a 1/100 dilution.
ICC/IF image of ab51504 stained A431 (Human epidermoid carcinoma cell line) cells.
The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51504, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
Anti-Cytokeratin 14 antibody [EP1612Y] (ab51054) at 1/20000 dilution + A431 (Human epidermoid carcinoma cell line) cell lysate at 10 µg
Goat anti-Rabbit-HRP at 1/2000 dilution
Predicted band size: 52 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?
Overlay histogram showing A431 (Human epidermoid carcinoma cell line) cells stained with ab51054 (red line).
The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab51054, 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 was rabbit IgG (monoclonal) ( 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.
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.
ab51054 has been referenced in 13 publications.
- Deng H et al. Establishment and optimization of epithelial cell cultures from human ectocervix, transformation zone, and endocervix optimization of epithelial cell cultures. J Cell Physiol 234:7683-7694 (2019). PubMed: 30609028
- Ragle LE et al. Long-label-retaining mammary epithelial cells are created early in ductal development and distributed throughout the branching ducts. Mech Dev 159:103565 (2019). PubMed: 31336167
- Mi B et al. The Designer Antimicrobial Peptide A-hBD-2 Facilitates Skin Wound Healing by Stimulating Keratinocyte Migration and Proliferation. Cell Physiol Biochem 51:647-663 (2018). PubMed: 30463067
- Lee JS et al. The Small Molecule NLRP3 Inflammasome Inhibitor MCC950 Does Not Alter Wound Healing in Obese Mice. Int J Mol Sci 19:N/A (2018). PubMed: 30360489
- Deng H et al. Susceptibility of epithelial cells cultured from different regions of human cervix to HPV16-induced immortalization. PLoS One 13:e0199761 (2018). PubMed: 29944714