Rabbit Recombinant Monoclonal Cytokeratin 14 antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
IHC-P | IP | WB | ICC/IF | Flow Cyt (Intra) | |
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Human | Tested | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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Species Human | Dilution info - | Notes - |
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Species Human | Dilution info - | Notes - |
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Species Human | Dilution info 1/100 | Notes - |
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Species Human | Dilution info - | Notes - |
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The 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.
Cytokeratin-14, Keratin-14, CK-14, K14, KRT14
Rabbit Recombinant Monoclonal Cytokeratin 14 antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ab243907 is the carrier-free version of Anti-Cytokeratin 14 antibody [EP1612Y] ab51054.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Cytokeratin 14 also known as CK14 keratin 14 or KRT14 is a type I keratin protein with a molecular mass of approximately 50 kDa. It forms intermediate filaments in epithelial cells primarily in the basal layer of stratified epithelium and is part of the cytoskeleton. Cytokeratin 14 expression appears in tissues such as the epidermis tongue and esophagus where it contributes to the structural integrity and mechanical stability of cells. Laboratories often utilize antibodies like anti-K14 and labels such as Alexa Fluor 555 or Alexa Fluor 647 to identify cytokeratin 14 in research settings.
This protein significantly contributes to the maintenance and regeneration of epithelial tissues by forming a network of filaments with keratin 5. KRT14 pairs with keratin 5 to create a keratin intermediate filament complex which plays a structural role in the resilience and elasticity of epithelial tissues. This partnership is essential for the assembly of the cytoskeleton and supports the protective barrier function of the skin and related tissues.
Cytokeratin 14 engages in pivotal cellular processes like keratinization and wound healing. The protein is actively involved in signal transduction pathways such as the epithelial cell differentiation pathway. Within these pathways KRT14 collaborates with KRT5 and other structural proteins to regulate cell proliferation and differentiation reflecting its importance in skin health and function.
Cytokeratin 14 associates with specific genetic conditions including epidermolysis bullosa simplex (EBS) and squamous cell carcinoma (SCC). In EBS mutations in the KRT14 gene lead to skin fragility. In SCC altered expression of KRT14 and related proteins such as keratin 5 can influence tumor progression and invasion. These associations highlight the critical role of KRT14 in both normal and pathological epithelial function.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054).
Immunohistochemical analysis of paraffin-embedded human squamous lung carcinoma tissue sections labeling Cytokeratin 14 with purified Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 at 1/100 dilution. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Sections were counterstained with Hematoxylin.
Antigen retrieval was heat mediated antigen retrieval using citrate buffer, pH 6.0).
This data was developed using Anti-Cytokeratin 14 antibody [EP1612Y] ab51054, the same antibody clone in a different buffer formulation.
Purified Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 at 1/20 dilution (0.5μg) immunoprecipitating Cytokeratin 14 in A431 whole cell lysate.
Lane 1 (input): A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+): Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 + A431 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 in A431 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (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: 48 kDa
All lanes: Immunoprecipitation - Anti-Cytokeratin 14 antibody [EP1612Y] (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054)
Predicted band size: 52 kDa
This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054). Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 staining KRT14 in wild-type A431 cells (top panel) and KRT14 knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then 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 Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 at 1/100 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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).
This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054).
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 observed at 49 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-Cytokeratin 14 antibody [EP1612Y] 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 Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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.
All lanes: Western blot - Anti-Cytokeratin 14 antibody [EP1612Y] (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054) at 1/10000 dilution
Lane 1: Wild-type A431 cell lysate at 20 µg
Lane 2: KRT14 knockout A431 cell lysate at 20 µg
Lane 2: Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (Human KRT14 (Cytokeratin 14) knockout A-431 cell line ab261897)
Lane 3: Human skin cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 49 kDa
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 Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054)
Overlay histogram showing A431 (Human epidermoid carcinoma cell line) cells stained with Anti-Cytokeratin 14 antibody [EP1612Y] 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 (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight®488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was was rabbit IgG (monoclonal) (1μg/1x106cells) 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 14 antibody [EP1612Y] ab51054).
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