PRODUCT CODE: ET1601-8

Cytokeratin 20 Recombinant Rabbit Monoclonal Antibody [SA35-03] (ET1601-8)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Rat

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Western blot analysis of Cytokeratin 20 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 (ET1601-8, 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: LOVO cell lysate<br />
Lane 2: HCT116 cell lysate<br />
Lane 3: CRC cell lysate
  • Western blot analysis of Cytokeratin 20 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 (ET1601-8, 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: LOVO cell lysate<br />
Lane 2: HCT116 cell lysate<br />
Lane 3: CRC cell lysate
  • ICC staining of Cytokeratin 20 in CRC 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 (ET1601-8, 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 rat small intestine tissue using anti-Cytokeratin 20 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 (ET1601-8, 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 colon carcinoma tissue using anti-Cytokeratin 20 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 (ET1601-8, 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 small intestine tissue using anti-Cytokeratin 20 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 (ET1601-8, 1/200) 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 20 was done on CRC cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-8, 1/50) (blue). 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; red).
Western blot analysis of Cytokeratin 20 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 (ET1601-8, 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: LOVO cell lysate
Lane 2: HCT116 cell lysate
Lane 3: CRC cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Cytokeratin 20 Recombinant Rabbit Monoclonal Antibody [SA35-03] (ET1601-8)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

LOVO cell lysate, HCT116 cell lysate, CRC cell lysate, CRC, rat small intestine tissue, human colon carcinoma tissue, human small intestine tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SA35-03

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

44 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:10-1:100

TARGET

UNIPROT #

PROTEIN NAME

Cytokeratin 20

SYNONYMS

CD20 antibody; CK 20 antibody; CK-20 antibody; CK20 antibody; Cytokeratin-20 antibody; Cytokeratin20 antibody; K1C20_HUMAN antibody; K20 antibody; KA20 antibody; Keratin 20 antibody; keratin 20, type I antibody; keratin 21, rat, homolog of antibody; Keratin antibody; Keratin type I cytoskeletal 20 antibody; Keratin-20 antibody; Keratin20 antibody; KRT 20 antibody; KRT 21 antibody; KRT20 antibody; KRT21 antibody; MGC35423 antibody; OTTHUMP00000164518 antibody; Protein IT antibody; type I cytoskeletal 20 antibody

SEQUENCE SIMILARITIES

Belongs to the intermediate filament family.

TISSUE SPECIFICITY

Expressed predominantly in the intestinal epithelium. Expressed in luminal cells of colonic mucosa. Also expressed in the Merkel cells of keratinized oral mucosa; specifically at the tips of some rete ridges of the gingival mucosa, in the basal layer of the palatal mucosa and in the taste buds of lingual mucosa.

DEVELOPMENTAL STAGE

First detected at embryonic week 8 in individual 'converted' simple epithelial cells of the developing intestinal mucosa. In later fetal stages, synthesis extends over most goblet cells and a variable number of villus enterocytes. In the developing gastric and intestinal mucosa, expressed in all enterocytes and goblet cells as well as certain 'low-differentiated' columnar cells, whereas the neuroendocrine and Paneth cells are negative.

POST-TRANSLATIONAL MODIFICATION

Hyperphosphorylation at Ser-13 occurs during the early stages of apoptosis but becomes less prominent during the later stages. Phosphorylation at Ser-13 also increases in response to stress brought on by cell injury (By similarity).; Proteolytically cleaved by caspases during apoptosis. Cleavage occurs at Asp-228.

SUBCELLULAR LOCATION

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, where they constitute up to 85% of mature keratinocytes in the vertebrate epidermis. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epi-thelial cells. The a-helical coiled-coil dimers associate laterally end-to-end to form 10 nm diameter filaments. Cytokeratins are useful markers of tissue differentiation, and in addition, they aid in the characterization of malignant tumors. Cytokeratin 20 is abundantly expressed in goblet cells and enterocytes of the gastrointestinal tract, and Cytokeratin 20 is a useful marker of pancreatic and colorectal cancer. Cytokeratin 20 is also helpful in distinguishing different types of highly related carcinomas, such as renal oncocytomas from renal cell carcinomas.