PRODUCT CODE: ET1608-32

Cytokeratin 8 Recombinant Rabbit Monoclonal Antibody [SU0338] (ET1608-32)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

Western blot analysis of Cytokeratin 8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-32, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: MCF-7 cell lysate<br />
Lane 3: A431 cell lysate
  • Western blot analysis of Cytokeratin 8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-32, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: MCF-7 cell lysate<br />
Lane 3: A431 cell lysate
  • ICC staining of Cytokeratin 8 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Cytokeratin 8 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Cytokeratin 8 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Cytokeratin 8 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Cytokeratin 8 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of Cytokeratin 8 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1608-32, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Cytokeratin 8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-32, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: MCF-7 cell lysate
Lane 3: A431 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Cytokeratin 8 Recombinant Rabbit Monoclonal Antibody [SU0338] (ET1608-32)

Immunogen

Synthetic peptide within human cytokeratin 8 aa 321-370 / 483.

Host

Rabbit

Positive Control

Hela cell lysate, MCF-7 cell lysate, A431 cell lysate, Hela, MCF-7, A431, human liver tissue, human breast carcinoma tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SU0338

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

53 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Cytokeratin 8

SYNONYMS

CARD2 antibody; Cell proliferation inducing gene 46 protein antibody; Cell proliferation inducing protein 46 antibody; CK 8 antibody; CK-8 antibody; CK8 antibody; CYK8 antibody; Cytokeratin 8 antibody; Cytokeratin-8 antibody; K2C8 antibody; K2C8_HUMAN antibody; K8 antibody; Keratin 8 antibody; Keratin antibody; keratin type II cytoskeletal 8 antibody; Keratin-8 antibody; KRT8 antibody; type II cytoskeletal 8 antibody; Type-II keratin Kb8 antibody

SEQUENCE SIMILARITIES

Belongs to the intermediate filament family.

TISSUE SPECIFICITY

Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.

POST-TRANSLATIONAL MODIFICATION

Phosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6.; Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.; O-GlcNAcylation increases solubility, and decreases stability by inducing proteasomal degradation.

SUBCELLULAR LOCATION

Nucleoplasm, Nucleus matrix, Cytoplasm.

FUNCTION

Cytokeratins comprise a diverse group of intermediate filament proteins (IFPs) that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. They have been found to be useful markers of tissue differentiation, which is directly applicable to the characterization of malignant tumors. Cytokeratin 8 expression is seen in epithelium and epithelium-derived tumors. The Cytokeratin 8 and 18 pair are normally expressed in simple epithelia, but not in stratified epithelial cells. Research indicates that squamous cell carcinomas derived from stratified epithelia show abnormal expression of Cytokeratin 8 and 18, although it is not known whether these proteins contribute to the malignant phenotype of the cells. Expression of Cytokeratin 8 and 18 in oral squamous cell carcinomas is an independent prognostic marker that indicates a poor prognosis. Cytokeratin 8 expression correlates with malignancy in leukoplakia and carcinomas of the head and neck; it is expressed in all non-small-cell lung cancers. Cytokeratin 8 has been shown to possess extracellular epitopes on tumor cells, which may represent valuable targets for therapy.