Anti-phospho-Keratin-8 Antibody (Ser23) (C15-1317-976)

General description

Keratin, type II cytoskeletal 8 (UniProt P05787; also known as CK-8, Cytokeratin-8, K8, Keratin-8, Type-II keratin Kb8) is encoded by the KRT8 (also known as CYK8) gene (Gene ID 3856) in human. Cytokeratin-8 (Keratin-8 or K8) belongs to a family of highly conserved epithelial-specific intermediate filament (IF) proteins that play an important role in the mechanical stability and integrity of epithelial cells and tissues. Each epithelium is characterized by its unique keratin expression pattern. Keratin 8 is a type II intermediate filament protein found in simple epithelia, where it pairs with keratin 18 (K8/KI8) and plays an important role in protecting cells from stress, injury, and apoptosis. The structure and function of K8/18 are regulated by posttranslational modifications, including glycosylation, ubiquitination, and phosphorylation. K8 has been reported to be phosphorylated on Ser8, Ser23, Ser73, and Ser431 by various protein kinases. PKCepsilon is reported to mediate K8 phosphorylation on Ser8 and Ser23, p38 and PKCdelta are responsible for K8 Ser73 phosphorylation, while ERK and JNK are involved in K8 Ser431 phosphorylation.

Specificity

This rabbit polyclonal antibody detected the target phospho band only in lysates from phosphatase inhibitors-treated, but not untreated HeLa cells. Antibody blocking by the immunogen peptide, but not the corresponding nonphosphorylated peptide prior to immunoblotting abolished target band detection. The Ser23 designation is based on human keratin-8 sequence with initiator Met1 removed. The phosphorylation site sequence in mouse/rat (-SSR(pS)YT-) differs from that of human (-SSR(pS)FT-) by a single Phe24-to-Tyr24 switch (F24Y). Cross-reactivity of this polyclonal antibody toward murine and rat species is likely, but has not been confirmed.

Immunogen

KLH-conjugated linear peptide corresponding to the phosphorylation site sequence containing the phosphorylated target serine.

Application

Peptide Inhibition Assay: Preincubation of a representative lot with the immunogen phosphopeptide, but not the corresponding nonphosphopeptide, abolished target band detection by Western blotting analysis of 10 µg lysate from phosphatase inhibitors-treated HeLa cells.

Research Category
Cell Structure

This rabbit polyclonal Anti-phospho-Keratin-8 (Ser23), Cat. No. ABT1372 is validated for use in Peptide Inhibition Assay and Western Blotting for the detection of Keratin 8 phosphorylated at serine 23.

Quality

Evaluated by Western Blotting in HeLa cells lysate.

Western Blotting Analysis: A 1:1,000 dilutiion of this antibody detected keratin-8 Ser23 phosphorylation in 10 µg of lysate from phosphatase inhibitors-treated, but not untreated, HeLa cells.

Target description

~54 kDa observed. 53.57/56.48 kDa (isoform 1/2) calculated (Met1 removed). Uncharacterized bands may be observed in some lysate(s).

Physical form

Affinity purified.

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

biological source: rabbit. Quality Level: 100. antibody form: affinity isolated antibody. antibody product type: primary antibodies. clone: polyclonal. purified by: affinity chromatography. species reactivity: human. species reactivity (predicted by homology): rhesus macaque (based on 100% sequence homology). technique(s): inhibition assay: suitable (peptide), western blot: suitable. NCBI accession no.: NP_002264. UniProt accession no.: P05787. shipped in: ambient. target post-translational modification: phosphorylation (pSer23). Gene Information: human ... KRT8(3856). Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.