KRT14 Human

Cytokeratin 14 Human Recombinant
Cat. No.
BT27584
Source
Escherichia Coli.
Synonyms
Keratin, type I cytoskeletal 14, Cytokeratin-14, CK-14, Keratin-14, K14, KRT14, NFJ, CK14, EBS3, EBS4.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

Cytokeratin 14 Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain having a molecular mass of 51,530 Dalton.
The KRT14 is purified by proprietary chromatographic techniques.

Product Specs

Introduction
Cytokeratin 14, a type I keratin, belongs to the keratin family, which constitutes the most diverse group of intermediate filaments. It typically forms a heterotetramer with two molecules of keratin 5, a type II keratin. This heterotetramer plays a crucial role in forming the cytoskeleton of epithelial cells. Mutations in the genes encoding these keratins are linked to a genetic condition called epidermolysis bullosa simplex. Notably, at least one pseudogene for cytokeratin 14 has been identified at the chromosomal location 17p12-p11.
Description
Recombinantly produced in E. coli, Cytokeratin 14 Human Recombinant is a single, non-glycosylated polypeptide chain with a molecular weight of 51,530 Daltons. The purification of KRT14 is achieved using proprietary chromatographic methods.
Physical Appearance
Sterile Filtered White lyophilized (freeze-dried) powder
Formulation
The protein was lyophilized at a concentration of 1 mg/ml from a sterile solution containing 30 mM Tris-HCl (pH 8), 9.5 M urea, 2 mM DTT, 2 mM EDTA, and 10 mM methylammonium chloride.
Solubility
To reconstitute the lyophilized CK-14, it is recommended to dissolve it in sterile 18 MΩ-cm H2O to a concentration of at least 100 µg/ml. This solution can then be further diluted in other aqueous solutions as needed.
Stability
While lyophilized KRT14 remains stable at room temperature for up to 3 weeks, it is recommended to store it desiccated below -18°C. Once reconstituted, KRT14 should be stored at 4°C for 2-7 days. For long-term storage, it is advisable to add a carrier protein such as HSA or BSA (0.1%) and store it below -18°C. Avoid repeated freeze-thaw cycles.
Purity
The purity of KRT14 is determined to be greater than 95.0% by SDS-PAGE analysis.
Reconstitution To Filaments
To reconstitute filaments, mix equimolar quantities of type I and type II cytokeratins, each at a concentration of approximately 0.5 mg/ml, dissolved in a 9.5 M urea buffer (as described above). Protofilaments and filament complexes can be obtained by dialyzing the polypeptide solution in a stepwise manner, first against a 4 M urea solution and then against a low salt solution (50 mM NaCl, 2 mM dithiothreitol, 10 mM Tris-HCl, pH 7.4). For immunization purposes, further dialysis against PBS (phosphate-buffered saline, such as Dulbecco's PBS) is recommended.
Synonyms
Keratin, type I cytoskeletal 14, Cytokeratin-14, CK-14, Keratin-14, K14, KRT14, NFJ, CK14, EBS3, EBS4.
Source
Escherichia Coli.

Product Science Overview

Structure and Function

Cytokeratin 14 is part of the keratin family, which consists of about 20 different cytokeratin isotypes in human cells . These proteins form intermediate filaments, which are essential components of the cytoskeleton. CK14 pairs with keratin 5 (KRT5) to form a heterodimer, which then assembles into keratin filaments . The nonhelical tail domain of CK14 is involved in promoting the self-organization of KRT5-KRT14 filaments into large bundles, enhancing the mechanical properties and resilience of keratin intermediate filaments .

Expression and Localization

CK14 is predominantly expressed in the basal compartment of all stratified squamous epithelia . This includes the epidermis, the lining of the oral cavity, esophagus, and the outer root sheath of hair follicles. It is also found in some glandular epithelia, myoepithelium, and mesothelial cells .

Clinical Significance

CK14 is a valuable marker in the diagnosis and differentiation of various epithelial tumors. It is particularly useful in distinguishing squamous cell carcinomas from poorly differentiated epithelial tumors . Additionally, CK14 is one of the specific basal markers for differentiating between basal and non-basal subtypes of breast carcinomas. It is also used to differentiate intraductal from invasive salivary duct carcinoma and to identify metaplastic carcinomas of the breast .

Recombinant CK14

Human recombinant CK14 is produced using recombinant DNA technology, which involves inserting the gene encoding CK14 into a suitable expression system, such as bacteria or mammalian cells. This allows for the production of large quantities of CK14 protein, which can be used for research and diagnostic purposes. Recombinant CK14 retains the same structural and functional properties as the native protein, making it a valuable tool in various applications, including immunohistochemistry, western blotting, and ELISA .

Applications

Recombinant CK14 is widely used in research to study the biology of epithelial cells and the role of keratins in cell structure and function. It is also used in the development of diagnostic assays for detecting CK14 expression in tissue samples. These assays are important for the diagnosis and classification of epithelial tumors, as well as for monitoring the progression of diseases involving epithelial cells .

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