Anti-KRT8 monoclonal antibody (DCABH-12162)

Mouse Anti-Human KRT8 monoclonal antibody for WB, IHC, IF, IP, FC

Specifications


Host Species
Mouse
Antibody Isotype
IgG1
Clone
L9/494
Species Reactivity
Human, Rat, Zebrafish
Immunogen
Cytoskeleton preparation containing cytokeratin 8.
Conjugate
Unconjugated

Applications


Application Notes
Flow Cytometry (0.5-1 ug/million cells);
Immunofluorescence (1-2 μg/mL);
Immunohistochemistry (1-2 μg/mL for 30 min at RT);
Immunoprecipitation (1-2 ug/500 ug protein);
WB (0.25-0.5 μg/mL) & The optimal working dilution should be determined by the end user.
*Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.

Target


Alternative Names
KRT8; keratin 8; keratin, type II cytoskeletal 8; CARD2; CK8; CYK8
Entrez Gene ID
UniProt ID

Product Background


Pathway
EGFR1 Signaling Pathway, organism-specific biosystem; Signaling mediated by p38-alpha and p38-beta, organism-specific biosystem;

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References


The Metastasis Potential Promoting Capacity of Cancer-Associated Fibroblasts Was Attenuated by Cisplatin via Modulating KRT8

ONCOTARGETS AND THERAPY

Authors: Li, Xueqin; Song, Qianqian; Guo, Xueru; Wang, Limin; Zhang, Qicheng; Cao, Limin; Ren, Yinghui; Wu, Xiang; Meng, Zhaowei; Xu, Ke

Background: Cancer-associated fibroblasts (CAFs) are an essential component of tumor microenvironment. They are attracting increasing attentions due to their crucial role in tumor growth, drug-resistance and metastasis. Cisplatin is a first-line chemotherapy drug applying in various types of cancer. There are intensive studies on cisplatin's effect on tumor cells, however, its effect on CAFs remains poorly understood. In the present study, we investigated the effect of cisplatin on CAFs. Methods: Cell migration was detected by wound healing assay. Cell invasion was performed by the transwell assay. mRNA expression was detected by quantitative PCR, and protein expression was detected by Western blotting. Tumor growth was measured using BALB/c nude mice tumor models. Results: Cisplatin attenuated the promoting capacity of CAFs on lung cancer cell migration and invasion, via suppressing CAFs' effect on metastasis-related genes including Twist1, vascular endothelial growth factor receptor (VEGFR), MMP2, and AKT signaling pathway. Keratin 8 (KRT8) was identified as a target of cisplatin. KRT8 upregulation in CAFs is responsible for the inhibitory effect of cisplatin on lung cancer cells metastasis potential through AKT pathway suppression. The stimulation of AKT by AKT activator SC79 reversed KRT8's effect on cell migration. Importantly, in vivo study also showed that CAFs enhanced tumor growth significantly, and cisplatin effectively abrogated the promoting effect of CAFs on tumor growth. Conclusion: Our results revealed a novel mechanism that cisplatin attenuated the metastasis promoting effect of CAFs via KRT8/AKT signaling pathway. This finding highlights KRT8 in CAFs as a potential therapeutic candidate for metastasis treatment.

Directed Expression of a Chimeric Type II Keratin Partially Rescues Keratin 5-null Mice

JOURNAL OF BIOLOGICAL CHEMISTRY

Authors: Alvarado, David M.; Coulombe, Pierre A.

The crucial role of structural support fulfilled by keratin intermediate filaments (IFs) in surface epithelia likely requires that they be organized into cross-linked networks. For IFs comprised of keratins 5 and 14 (K5 and K14), which occur in basal keratinocytes of the epidermis, formation of cross-linked bundles is, in part, self-driven through cis-acting determinants. Here, we targeted the expression of a bundling-competent KRT5/KRT8 chimeric cDNA (KRT8bc) or bundling-deficient wild type KRT8 as a control to the epidermal basal layer of Krt5-null mice to assess the functional importance of keratin IF self-organization in vivo. Such targeted expression of K8bc rescued Krt5-null mice with a 47% frequency, whereas K8 completely failed to do so. This outcome correlated with lower than expected levels of K8bc and especially K8 mRNA and protein in the epidermis of E18.5 replacement embryos. Ex vivo culture of embryonic skin keratinocytes confirmed the ability of K8bc to form IFs in the absence of K5. Additionally, electron microscopy analysis of E18.5 embryonic skin revealed that the striking defects observed in keratin IF bundling, cytoarchitecture, and mitochondria are partially restored by K8bc expression. As young adults, viable KRT8bc replacement mice develop alopecia and chronic skin lesions, indicating that the skin epithelia are not completely normal. These findings are consistent with a contribution of self-mediated organization of keratin IFs to structural support and cytoarchitecture in basal layer keratinocytes of the epidermis and underscore the importance of context-dependent regulation for keratin genes and proteins in vivo.

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