Cytokeratin 6 Recombinant Rabbit Monoclonal Antibody [SN71-07]
cat.: ET1611-70
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SN71-07
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: 60 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Cytokeratin 6 aa 1-118 / 564.
Positive control: A431 cell lysate, human skin tissue lysate, A431, rat skin tissue, human tonsil tissue.
Subcellular location: Keratin filament, cytosol, extracellular exosome, nucleus, membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:500-1:2,000
1:100-1:500
1:100-1:500
1:50-1:200
Uniprot #: SwissProt: P02538 Human | P04259 Human | P48668 Human | Q4FZU2 Rat
Alternative names: CK6 CK 6A CK 6B CK 6C CK 6D CK 6E CK-6B CK-6C CK-6E Cytokeratin 6a Cytokeratin 6B Cytokeratin 6C Cytokeratin 6D Cytokeratin 6E Cytokeratin-6B Cytokeratin-6C Cytokeratin-6E K2C6C_HUMAN K6a keratin K6b keratin K6C K6c keratin K6d keratin K6e keratin Keratin Keratin K6h Keratin type II cytoskeletal 6A Keratin type II cytoskeletal 6B Keratin type II cytoskeletal 6C Keratin type II cytoskeletal 6D Keratin type II cytoskeletal 6E Keratin-6C KRT6A KRT6B KRT6C KRT6D KRT6E type II cytoskeletal 6C Type-II keratin Kb12
Images
ET1611-70_1.jpg Fig1: Western blot analysis of Cytokeratin 6 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 (ET1611-70, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: A431 cell lysate
Lane 2: Human skin tissue lysate
ET1611-70_2.jpg Fig2: ICC staining of Cytokeratin 6 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 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-70, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1611-70_3.jpg Fig3: Immunofluorescence analysis of paraffin-embedded rat skin tissue labeling Cytokeratin 6 (ET1611-70).

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 6 (ET1611-70, red) at 1/50 dilution at +4℃ overnight, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibodies at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1611-70_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Cytokeratin 6 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-70, 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.
ET1611-70_5.jpg Fig5: Western blot analysis of Cytokeratin 6 on different lysates with Rabbit anti-Cytokeratin 6 antibody (ET1611-70) at 1/1,000 dilution.

Lane 1: HaCaT cell lysate
Lane 2: A431 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Exposure time: 10 seconds; ECL: K1801;

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 (ET1611-70) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1611-70_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Cytokeratin 6 antibody (ET1611-70) at 1/100 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 (ET1611-70) at 1/100 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.
ET1611-70_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma tissue with Rabbit anti-Cytokeratin 6 antibody (ET1611-70) at 1/100 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 (ET1611-70) at 1/100 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.
ET1611-70_8.jpg Fig8: Immunocytochemistry analysis of A431 cells labeling Cytokeratin 6 with Rabbit anti-Cytokeratin 6 antibody (ET1611-70) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 6 antibody (ET1611-70) 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.
ET1611-70_9.jpg Fig9: Flow cytometric analysis of A431 cells labeling Cytokeratin 6.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-70, 1μg/mL) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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