Product Name
Cytokeratin 4 Recombinant Rabbit Monoclonal Antibody [SN74-03] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human Cytokeratin 4 aa 395-520 / 520.
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 56 kDa
Positive Control
A431, human tonsil tissue, human esophagus tissue.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
Cytokeratins are a subfamily of intermediate filament keratins that are characterized by a remarkable biochemical diversity, which is represented in human epithelial tissues by at least 20 different polypeptides. Cytokeratins range in isoelectric range between 4.9 and 7.8. Cytokeratin 1 has the highest molecular weight, while Cytokeratin 19 has the lowest molecular weight. The cytokeratins are divided into the type I and type II subgroups. Type II family members comprise the basic to neutral members, Cytokeratins 1-8, while the type I group comprises the acidic members, Cytokeratins 9-20. Various epithelia in the human body usually express cytokeratins which are characteristic of the type of epithelium and related to the degree of maturation or differentiation within said epithelium. Cytokeratin subtype expression patterns are used to an increasing extent in the distinction of different types of epithelial malignancies. Cytokeratin 4 is expressed in differentiated layers of the mucosal and esophageal epithelia along with Cytokeratin 13.
Background References
1. Jacot TA et al. Development of a composite measure of product adherence, protocol compliance, and semen exposure using DNA and protein biomarkers for topical HIV prevention studies. PLoS One 9:e114368 (2014).
2. Badylak SF et al. Esophageal preservation in five male patients after endoscopic inner-layer circumferential resection in the setting of superficial cancer: a regenerative medicine approach with a biologic scaffold. Tissue Eng Part A 17:1643-50 (2011).
Sequence Similarity
Belongs to the intermediate filament family.
Tissue Specificity
Detected in the suprabasal layer of the stratified epithelium of the esophagus, exocervix, vagina, mouth and lingual mucosa, and in cells and cell clusters in the mucosa and serous gland ducts of the esophageal submucosa (at protein level). Expressed widely in the exocervix and esophageal epithelium, with lowest levels detected in the basal cell layer.
Subcellular Location
Cell surface, intermediate filament cytoskeleton, nucleus.
Synonyms
CK 4 antibody
CK-4 antibody
CK4 antibody
CYK4 antibody
Cytokeratin 4 antibody
Cytokeratin-4 antibody
Cytokeratin4 antibody
FLJ31692 antibody
K2C4_HUMAN antibody
K4 antibody
Expand
CK 4 antibody
CK-4 antibody
CK4 antibody
CYK4 antibody
Cytokeratin 4 antibody
Cytokeratin-4 antibody
Cytokeratin4 antibody
FLJ31692 antibody
K2C4_HUMAN antibody
K4 antibody
Keratin 4 antibody
Keratin antibody
Keratin type II cytoskeletal 4 antibody
Keratin-4 antibody
Keratin4 antibody
KRT 4 antibody
Krt4 antibody
type II cytoskeletal 4 antibody
Type-II keratin Kb4 antibody
Collapse
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This data was developed using ET1611-71, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of A431 cells labeling Cytokeratin 4 with Rabbit anti-Cytokeratin 4 antibody (ET1611-71) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cytokeratin 4 antibody (ET1611-71) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using ET1611-71, the same antibody clone in a different buffer formulation.
Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling Cytokeratin 4 (ET1611-71) and Vimentin (EM0401).
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Cytokeratin 4 (ET1611-71, red) at 1/200 dilution and Vimentin (EM0401, green) at 1/200 dilution at +4℃ overnight, washed with PBS.
Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) and Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) were used as the secondary antibodies at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
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This data was developed using ET1611-71, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-Cytokeratin 4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-71, 1/400) 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.
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This data was developed using ET1611-71, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Cytokeratin 4 antibody.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-71, 1/400) 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.
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This data was developed using ET1611-71, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of Cytokeratin 4 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-71, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-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).
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