WB
Western blot analysis of pan Cytokeratin on different lysates with Mouse anti-pan Cytokeratin antibody at 1/2,000 dilution. Lane 1: A431 cell lysate (10 µg/Lane), Lane 2: HeLa cell lysate (10 µg/Lane), Lane 3: MCF7 cell lysate (10 µg/Lane), Lane 4: SK-Br-3 cell lysate (10 µg/Lane), Lane 5: MDA-MB-231 cell lysate (10 µg/Lane), Lane 6: MDA-MB-468 cell lysate (10 µg/Lane), Lane 7: Human liver tissue lysate (20 µg/Lane), Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-pan Cytokeratin antibody at 1/2,000 dilution. 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.IHC
Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Mouse anti-pan Cytokeratin antibody at 1/2,000 dilution. 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.IHC
Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Mouse anti-pan Cytokeratin antibody at 1/1,000 dilution. 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour 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.ICC/IF
Immunocytochemistry analysis of A431 cells labeling pan Cytokeratin with Mouse anti-pan Cytokeratin antibody 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 Mouse anti-pan Cytokeratin antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.IF-P
Immunofluorescence analysis of paraffin-embedded human stomach cancer tissue labeling pan Cytokeratin with Mouse anti-pan Cytokeratin antibody at 1/200 dilution. 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 antibody ( green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).FC
Flow cytometric analysis of A431 cells labeling pan Cytokeratin. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | pan Cytokeratin Recombinant Mouse Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Full length native protein corresponding to Human pan Cytokeratin. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG1 |
| Host Species | Mouse |
| Tested Applications | FCICC/IFIF-PIHCWB |
| WB:1:2000 IHC:1:1000-1:2000 ICC/IF:1:100 IF-P:1:200 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | pan Cytokeratin antibody pan-Cytokeratin antibody pan ck antibody pan-ck antibody panck antibody pan Keratin antibody |
|---|---|
| Molecular Weight(MW) | 27kDa |
| 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. Cytokeratins have been found to be useful markers of tissue differentiation which is directly applicable to the characterization of malignant tumors. For example, cytokeratins 10 and 13 are expressed highly in a subset of squamous cell carcinomas while cytokeratin 18 is expressed in a majority of adenocarcinomas and basal cell carcinomas. Since each epithelium contains at least one acidic and one basic keratin, this antibody is used to observe the distribution of keratin-containing cells in normal epithelia and to identify neoplasms derived from such epithelium. Recognizes humancytokeratins 10, 14, 15, 16 and 19, and cytokeratins 1, 2, 3, 4, 5, 6, 7 and 8. |
| Cellular Localization | Cytoplasmic |
WB
Western blot analysis of pan Cytokeratin on different lysates with Mouse anti-pan Cytokeratin antibody at 1/2,000 dilution. Lane 1: A431 cell lysate (10 µg/Lane), Lane 2: HeLa cell lysate (10 µg/Lane), Lane 3: MCF7 cell lysate (10 µg/Lane), Lane 4: SK-Br-3 cell lysate (10 µg/Lane), Lane 5: MDA-MB-231 cell lysate (10 µg/Lane), Lane 6: MDA-MB-468 cell lysate (10 µg/Lane), Lane 7: Human liver tissue lysate (20 µg/Lane), Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-pan Cytokeratin antibody at 1/2,000 dilution. 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.
IHC
Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Mouse anti-pan Cytokeratin antibody at 1/2,000 dilution. 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour 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.
IHC
Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue with Mouse anti-pan Cytokeratin antibody at 1/1,000 dilution. 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour 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.
ICC/IF
Immunocytochemistry analysis of A431 cells labeling pan Cytokeratin with Mouse anti-pan Cytokeratin antibody 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 Mouse anti-pan Cytokeratin antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.
IF-P
Immunofluorescence analysis of paraffin-embedded human stomach cancer tissue labeling pan Cytokeratin with Mouse anti-pan Cytokeratin antibody at 1/200 dilution. 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 antibody ( green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
FC
Flow cytometric analysis of A431 cells labeling pan Cytokeratin. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:2000 IHC:1:1000-1:2000 ICC/IF:1:100 IF-P:1:200 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
