antibody

Anti-Cytokeratin 8 Antibody [SU0338]

ET1608-32TR

20 uL

99 USD

monoclonal

domestic rabbit

nonconjugated

SU0338

human, mouse

western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

Anti-Cytokeratin 8 Antibody [SU0338]

Human,Mouse

WB,IF-Cell,IF-Tissue,IHC-P,IP,FC

Non-conjugated

Synthetic peptide within Human Cytokeratin 8 aa 321-370 / 483.

SwissProt: P05787 Human;SwissProt: P11679 Mouse

Rabbit

SU0338

IgG

20 uL

99 USD

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Liquid

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Protein A affinity purified.

Recombinant Rabbit monoclonal Antibody

Hela cell lysate, A431 cell lysate, Hela, MCF-7, A431, human liver tissue, human breast carcinoma tissue, human breast tissue, SK-Br-3, HepG2.

Predicted band size: 54 kDa

Nucleoplasm, Nucleus matrix, Cytoplasm.

1 mg/mL.

WB: 1:1,000-1:2,000 ;IF-Cell: 1:400-1:800 ;IF-Tissue: 1:400-1:800 ;IHC-P: 1:50-1:1,500 ;FC: 1:500-1:1,000 ;IP: Use at an assay dependent concentration.

ET1608-32_2.jpg

Western blot analysis of Cytokeratin 8 on different lysates with Rabbit anti-Cytokeratin 8 antibody (ET1608-32) at 1/2,000 dilution. Lane 1: Hela-si NT cell lysate Lane 2: Hela-si Cytokeratin 8 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 54 kDa Observed band size: 54 kDa Exposure time: 24 seconds; 4-20% SDS-PAGE gel. ET1608-32 was shown to specifically react with Cytokeratin 8 in Hela-si NT cells. Weakened band was observed when Hela-si Cytokeratin 8 sample was tested. Hela-si NT and Hela-si Cytokeratin 8 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1608-32, 1/2,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were 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.

ET1608-32_3.jpg

ICC staining of Cytokeratin 8 in Hela 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 (ET1608-32, 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).

ET1608-32_4.jpg

ICC staining of Cytokeratin 8 in MCF-7 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 (ET1608-32, 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).

ET1608-32_5.jpg

ICC staining of Cytokeratin 8 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 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-32, 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).

ET1608-32_6.jpg

Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Cytokeratin 8 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 (ET1608-32, 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.

ET1608-32_7.jpg

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Cytokeratin 8 antibody (ET1608-32) 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 (ET1608-32) 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.

ET1608-32_8.jpg

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Cytokeratin 8 antibody (ET1608-32) 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 (ET1608-32) 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.

ET1608-32_9.jpg

Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Cytokeratin 8 antibody (ET1608-32) 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 (ET1608-32) 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.

ET1608-32_10.jpg

Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 8 (ET1608-32). 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 8 (ET1608-32, red) at 1/400 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).

ET1608-32_11.jpg

Immunofluorescence analysis of paraffin-embedded human liver tissue labeling Cytokeratin 8 (ET1608-32) 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 8 (ET1608-32, red) at 1/50 dilution and Vimentin (EM0401, green) at 1/500 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).

ET1608-32_12.jpg

Immunocytochemistry analysis of SK-Br-3 cells labeling Cytokeratin 8 (ET1608-32). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.05% Triton X-100 in PBS for 10 minutes, and then blocked with 2% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody Cytokeratin 8 (ET1608-32, red) at 1/50 dilution and Beta-tubulin (EM0103, yellow) at 1/500 dilution at +4℃ overnight. 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).

ET1608-32_13.jpg

Flow cytometric analysis of HepG2 cells labeling Cytokeratin 8. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1608-32, 1ug/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).

ET1608-32_14.jpg

Cytokeratin 8 was immunoprecipitated in 0.2mg HeLa cell lysate with ET1608-32 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1608-32 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: Rabbit IgG instead of ET1608-32 in HeLa cell lysate Lane 3: ET1608-32 IP in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 43 seconds