KRT14 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 13 bp deletion Frameshift = 99.9%.
Images
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 13 bp deletion Frameshift = 99.9%
Alternative names
CK-14, Cytokeratin-14, Dowling Meara, EBS3, EBS4, Epidermolysis bullosa simplex, K14, K1C14_HUMAN, Keratin, Keratin 14 (epidermolysis bullosa simplex, Dowling-Meara, Koebner), Keratin type I cytoskeletal 14, Keratin-14, Koebner, Krt 14, NFJ, OTTHUMP00000164624, type I cytoskeletal 14
Recommended products
KRT14 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 13 bp deletion Frameshift = 99.9%.
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 13 bp deletion Frameshift = 99.9%
- Disease
- Epidermoid Carcinoma
- Concentration
Properties
- Gene name
- KRT14
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Biological function summary
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.
- Pathways
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.
- Associated diseases and disorders
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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10 product images
Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Cytokeratin 14 antibody [EPR17336] ab197893 observed at 52 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.Anti-Cytokeratin 14 antibody [EPR17336] ab197893 was shown to react with KRT14 in A431 wild-type cells in Western blot. Loss of signal was observed when KRT14 knockout sample was used. A431 wild-type 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 [EPR17336] ab197893 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 50000 dilution and a 1 in 20000 dilution respectively. Blots were developed 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 [EPR17336] (Anti-Cytokeratin 14 antibody [EPR17336] ab197893) at 1/50000 dilution
Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2: KRT14 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Lane 3: Human skin whole tissue 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: 52 kDa
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Alexa Fluor® 647 Anti-Cytokeratin 14 antibody [EP1612Y] ab192056 staining KRT14 in wild-type A-431 cells (top panel) and KRT14 knockout A-431 cells (ab261897) (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 Alexa Fluor® 647 Anti-Cytokeratin 14 antibody [EP1612Y] ab192056 at 1/100 dilution and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887 (Mouse monoclonal to alpha Tubulin - Alexa Fluor® 488) at 1/250 dilution overnight at 4oC. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] ab192055 staining KRT14 in wild-type A-431 cells (top panel) and KRT14 knockout A-431 cells (ab261897) (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 Alexa Fluor® 488 Anti-Cytokeratin 14 antibody [EP1612Y] ab192055 at 1/100 dilution and Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab190573 (Rabbit monoclonal to alpha Tubulin - Alexa Fluor® 647) at 1/250 dilution overnight at 4oC. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Next Generation Sequencing - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Knockout achieved by CRISPR/Cas9; X = 13 bp deletion; Frameshift = 99.9%
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Anti-Cytokeratin 14 antibody [EP1612Y] ab51054 staining KRT14 in wild-type A-431 cells (top panel) and KRT14 knockout A-431 cells (ab261897) (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 4oC 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 ug/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 ug/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Anti-Cytokeratin 14 antibody [SP53] ab119695 staining KRT14 in wild-type A-431 cells (top panel) and KRT14 knockout A-431 cells (ab261897) (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 [SP53] ab119695 at 5?g/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4oC 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 ?g/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 ?g/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
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° 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 (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
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Anti-Cytokeratin 14 antibody [LL002] ab7800 staining KRT14 in wild-type A-431 cells (top panel) and KRT14 knockout A-431 cells (ab261897) (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 [LL002] ab7800 at 1ug/ml concentration and Anti-beta Tubulin antibody - Loading Control ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4oC followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 ug/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 2 ug/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Anti-Cytokeratin 14 antibody [EPR1612] ab108417 staining KRT14 in wild-type A-431 cells (top panel) and KRT14 knockout A-431 cells (ab261897) (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 [EPR1612] ab108417 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).
Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (ab261897)
Anti-Cytokeratin 14 antibody [EPR17350] - Cytoskeleton Marker ab181595 staining Cytokeratin 14 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 [EPR17350] - Cytoskeleton Marker ab181595 at 0.2μg/ml concentration 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).
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