AB261897

Human KRT14 (Cytokeratin 14) knockout A-431 cell line

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KRT14 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 13 bp deletion Frameshift = 99.9%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

11 Images

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab108417 at 1/100 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).

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab51054 at 1/100 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4^oC followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 ug/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (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).

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab7800 at 1ug/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4^oC followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 ug/ml (shown in green) and a goat secondary antibody to rabbit IgG (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).

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab181595 at 0.2μg/ml concentration 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).

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab119695 at 5 µg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4^oC 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).

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab192055 at 1/100 dilution and ab190573 (Rabbit monoclonal to alpha Tubulin - Alexa Fluor^® 647) at 1/250 dilution overnight at 4^oC. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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 ab192056 at 1/100 dilution and ab195887 (Mouse monoclonal to alpha Tubulin - Alexa Fluor^® 488) at 1/250 dilution overnight at 4^oC. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Lab

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 2 : KRT14 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 3 : Human skin whole tissue lysate 20 ug

Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab119695 observed at 52 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab119695 was shown to react with KRT14 in wild-type A-431 cells in Western blot Loss of signal was observed when KRT14 knockout cell line ab261897 (knockout cell lysate ab261706) was used. Wild-type A-431 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 ab119695 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 93 dilution and a 1 in 20000 dilution respectively. Blots were developed 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.

All lanes:

Western blot - Anti-Cytokeratin 14 antibody [SP53] (<a href='/en-us/products/primary-antibodies/cytokeratin-14-antibody-sp53-ab119695'>ab119695</a>) at 1/93 dilution

Lane 1:

Wild-type 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 lysate (<a href='/en-us/products/cell-lysates/human-krt14-cytokeratin-14-knockout-a-431-cell-lysate-ab261706'>ab261706</a>) at 20 µg

Lane 3:

Human skin whole tissue lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 52 kDa,37 kDa

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Lab

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Lanes 1 - 4 : Merged signal (red and green). Green - ab197893 observed at 52 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

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 ab197893 and 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 (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.

All lanes:

Western blot - Anti-Cytokeratin 14 antibody [EPR17336] (<a href='/en-us/products/primary-antibodies/cytokeratin-14-antibody-epr17336-ab197893'>ab197893</a>) 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

Predicted band size: 52 kDa

Observed band size: 52 kDa

false

Lab

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

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° 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.

All lanes:

Western blot - Anti-Cytokeratin 14 antibody [EP1612Y] (<a href='/en-us/products/primary-antibodies/cytokeratin-14-antibody-ep1612y-ab51054'>ab51054</a>) 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

Predicted band size: 52 kDa

Observed band size: 49 kDa

false

Supplier Data

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%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Form

Liquid

form

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "ICC/IF": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

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Product details

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.

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What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab261897-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab261897 Human KRT14 (Cytokeratin 14) knockout A-431 cell line", "number":"AB261897-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab263975 Human wild-type A-431 cell line", "number":"AB261897-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab261897-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab261897 Human KRT14 (Cytokeratin 14) knockout A-431 cell line", "number":"AB261897-CMP01", "productcode":"" } ] } } }

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Properties and storage information

Gene name

KRT14

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Immunocytochemistry, Next Generation Sequencing, Western blot

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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.

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human KRT14 (Cytokeratin 14) knockout A-431 cell line

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Immunocytochemistry/ Immunofluorescence - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Next Generation Sequencing - Human KRT14 (Cytokeratin 14) knockout A-431 cell line (AB261897)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Disease

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Initial handling guidelines

Subculture guidelines

Culture medium

Cryopreservation medium

Product protocols

Product promise