Cytokeratin 4 Recombinant Rabbit Monoclonal Antibody [SN74-03]
cat.: ET1611-71
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | SN74-03 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 56 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human Cytokeratin 4 aa 395-520 / 520. |
| Positive control: | A431, human tonsil tissue, human esophagus tissue. |
| Subcellular location: | Cell surface, intermediate filament cytoskeleton, nucleus. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P FC IP |
1:500-1:1,000 1:50-1:200 1:50-1:200 1:100-1:500 1:50-1:100 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: P19013 Human |
| Alternative names: | CK 4 CK-4 CK4 CYK4 Cytokeratin 4 Cytokeratin-4 Cytokeratin4 FLJ31692 K2C4_HUMAN K4 Keratin 4 Keratin Keratin type II cytoskeletal 4 Keratin-4 Keratin4 KRT 4 Krt4 type II cytoskeletal 4 Type-II keratin Kb4 |
Images
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Fig1:
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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Fig2:
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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Fig3: 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) for 20 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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Fig4:
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) for 20 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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Fig5: 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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.