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AB261706

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

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KRT14 KO cell lysate available now. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 13 bp deletion Frameshift = 99.9%.

View 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

4 Images
Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)
  • WB

Lab

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)

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 - 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 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 ab197893 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C 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 (<a href='/en-us/products/cell-lines/human-krt14-cytokeratin-14-knockout-a-431-cell-line-ab261897'>ab261897</a>)

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

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)
  • WB

Lab

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : KRT14 knockout A431 cell lysate 20 ug

Lane 3 : Human skin cell 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 - 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 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 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.

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 (<a href='/en-us/products/cell-lines/human-krt14-cytokeratin-14-knockout-a-431-cell-line-ab261897'>ab261897</a>)

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

Next Generation Sequencing - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)
  • NGS

Supplier Data

Next Generation Sequencing - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)

Knockout achieved by CRISPR/Cas9; X = 13 bp deletion; Frameshift = 99.9%

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)
  • WB

Lab

Western blot - Human KRT14 (Cytokeratin 14) knockout A-431 cell lysate (AB261706)

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 (ab261706) 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

false

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

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

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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?

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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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°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

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Product protocols

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