Cytokeratin 14 Recombinant Rabbit Monoclonal Antibody [SC65-06]
cat.: ET1610-42
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: SC65-06
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: 51 kDa
Isotype: IgG
Immunogen: Recombinant protein within Mouse Cytokeratin 14 aa 250-484 / 484.
Positive control: A431 cell lysate, mouse skin tissue lysate, B16F1, SW480, HepG2, human cervical carcinoma tissue, human skin tissue, rat skin tissue, mouse skin tissue, mouse prostate tissue.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:2,000
1:100-1:500
1:100-1:500
1:100-1:1,500
1:1,000
Uniprot #: SwissProt: P02533 Human | Q61781 Mouse | Q6IFV1 Rat
Alternative names: CK 14 CK-14 ck14 Cytokeratin 14 Cytokeratin-14 Cytokeratin14 Dowling Meara EBS3 EBS4 Epidermolysis bullosa simplex K14 K1C14_HUMAN Keratin 14 (epidermolysis bullosa simplex, Dowling-Meara, Koebner) Keratin 14 Keratin Keratin type I cytoskeletal 14 Keratin, type I cytoskeletal 14 Keratin-14 Keratin14 Koebner Krt 14 Krt14 NFJ OTTHUMP00000164624 type I cytoskeletal 14
Images
ET1610-42_1.jpg Fig1: Western blot analysis of Cytokeratin 14 on different lysates with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) at 1/2,000 dilution.

Lane 1: A431 cell lysate (15 µg/Lane)
Lane 2: PANC-1 cell lysate (15 µg/Lane) (negative)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane) (negative)
Lane 4: PC-12 cell lysate (15 µg/Lane) (negative)
Lane 5: Mouse skin tissue lysate (20 µg/Lane)

Predicted band size: 51 kDa
Observed band size: 51 kDa

Exposure time: 24 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 (ET1610-42) at 1/2,000 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.
ET1610-42_2.jpg Fig2: ICC staining of Cytokeratin 14 in B16F1 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 (ET1610-42, 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).
ET1610-42_3.jpg Fig3: ICC staining of Cytokeratin 14 in SW480 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 (ET1610-42, 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).
ET1610-42_4.jpg Fig4: ICC staining of Cytokeratin 14 in HepG2 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 (ET1610-42, 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).
ET1610-42_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) 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 (ET1610-42) 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.
ET1610-42_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) at 1/1,500 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 (ET1610-42) at 1/1,500 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.
ET1610-42_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) at 1/1,500 dilution.

The section was not undergone antigen retrieval. 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 (ET1610-42) at 1/1,500 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.
ET1610-42_8.jpg Fig8: Immunofluorescence analysis of paraffin-embedded rat skin tissue labeling Cytokeratin 14 (ET1610-42).

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 14 (ET1610-42, red) at 1/500 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).
ET1610-42_9.jpg Fig9: Immunofluorescence analysis of paraffin-embedded rat skin tissue labeling Cytokeratin 14 (ET1610-42) 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 14 (ET1610-42, red) at 1/400 dilution and Vimentin (EM0401, green) at 1/400 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).
ET1610-42_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-Cytokeratin 14 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 (ET1610-42, 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.
ET1610-42_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded mouse prostate tissue using anti-Cytokeratin 14 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 (ET1610-42, 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.
ET1610-42_12.jpg Fig12: Immunocytochemistry analysis of A431 cells labeling Cytokeratin 14 with Rabbit anti-Cytokeratin 14 antibody (ET1610-42) at 1/500 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 14 antibody (ET1610-42) at 1/500 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.
ET1610-42_13.jpg Fig13: Flow cytometric analysis of A431 cells labeling Cytokeratin 14.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-42, 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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