KRT8 Human

Cytokeratin 8 Human Recombinant
Cat. No.
BT28567
Source
Escherichia Coli.
Synonyms
Keratin type II cytoskeletal 8, Cytokeratin-8, CK-8, Keratin-8, K8, KRT8, CYK8, KO, CK8, K2C8, CARD2.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis 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 8 Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain having a molecular mass of 53,532 Dalton.
The KRT8 is purified by proprietary chromatographic techniques.

Product Specs

Introduction
Keratin 8 and 18 (K8/18) are the primary intermediate filament (IF) proteins found in simple epithelia, which are composed of a single layer of cells.
Description
Recombinant human Cytokeratin 8, expressed in E. coli, is a non-glycosylated polypeptide chain with a molecular weight of 53,532 Daltons. The purification of KRT8 is achieved through proprietary chromatographic techniques.
Physical Appearance
Sterile white powder obtained by lyophilization (freeze-drying).
Formulation
The protein solution (1 mg/ml) was lyophilized 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 KRT8, it is recommended to dissolve it in sterile 18 MΩ·cm H2O at a concentration of at least 100 µg/ml. This solution can then be further diluted with other aqueous solutions as needed.
Stability
Lyophilized KRT8 remains stable at room temperature for 3 weeks; however, it is recommended to store it at 2-8°C. After reconstitution, KRT8 should be stored at 4°C for 2-7 days. For long-term storage, it is advised to store it below -18°C. The addition of a carrier protein (0.1% HSA or BSA) is recommended for long-term storage. Avoid repeated freeze-thaw cycles.
Purity
The purity is determined to be greater than 95.0% using the following methods: (a) RP-HPLC analysis, (b) SDS-PAGE analysis.
Reconstitution To Filaments
To form filaments, mix equal molar amounts of type I and type II cytokeratins at a concentration of approximately 0.5 mg/ml, both dissolved in a 9.5 M urea buffer (see above). Protofilaments and filament complexes can be obtained by stepwise dialysis of the polypeptide solution against a 4 M urea solution and then against a low salt buffer (50 mM NaCl, 2 mM dithiothreitol, 10 mM Tris-HCl, pH 7.4). For immunization purposes, the solution can be further dialyzed against PBS (e.g., Dulbecco's PBS).
Synonyms
Keratin type II cytoskeletal 8, Cytokeratin-8, CK-8, Keratin-8, K8, KRT8, CYK8, KO, CK8, K2C8, CARD2.
Source
Escherichia Coli.

Product Science Overview

Introduction

Cytokeratin 8 (CK8), also known as Keratin 8 (K8), is a type II intermediate filament protein that plays a crucial role in the structural integrity and function of epithelial cells. It is encoded by the KRT8 gene located on the long arm of chromosome 12 . CK8 is typically found in simple single-layered epithelial cells and is known to heteropolymerize with Keratin 18 (K18) to form intermediate filaments .

Structure and Properties

Recombinant Human Cytokeratin 8 (rHuCTK-8) is produced in Escherichia coli (E. coli) and is a single, non-glycosylated polypeptide chain with a molecular mass of approximately 53,532 Daltons . The recombinant protein is purified using proprietary chromatographic techniques to ensure high purity and quality . The amino acid sequence of CK8 includes several key regions that contribute to its structural and functional properties .

Biological Function

CK8, along with K18, forms a network of intermediate filaments that provide mechanical support to epithelial cells. This network is essential for maintaining cell shape, integrity, and resilience against mechanical stress. CK8 is also involved in various cellular processes, including apoptosis, cell signaling, and differentiation .

Applications

Recombinant CK8 is widely used in research to study the function and regulation of intermediate filaments in epithelial cells. It is also utilized in various assays, including ELISA, immunoaffinity purification, protein arrays, and Western blotting . The recombinant protein’s high purity and stability make it a valuable tool for investigating the molecular mechanisms underlying epithelial cell biology.

Storage and Handling

Recombinant CK8 should be stored at -80°C to maintain its stability and prevent degradation. It is recommended to avoid repeated freeze-thaw cycles to preserve the protein’s functionality . The protein is typically supplied in a buffer containing Tris-HCl and reduced glutathione, which helps maintain its structural integrity .

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