WB
Western blot analysis of Cytokeratin 20 on different lysates with Rabbit anti-Cytokeratin 20 antibody at 1/5,000 dilution. Lane 1: HT-29 cell lysate, Lane 2: LoVo cell lysate, Lane 2: CRC cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 1 minute; 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 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit 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 Rabbit anti-Cytokeratin 20 antibody at 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 at 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.IHC
Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-Cytokeratin 20 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 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
ICC staining of Cytokeratin 20 in CRC 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 (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).FC
Flow cytometric analysis of CRC cells with Cytokeratin 20 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.IP
Cytokeratin 20 was immunoprecipitated from 0.2 mg HT-29 cell lysate with Rabbit anti-Cytokeratin 20 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Cytokeratin 20 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HT-29 cell lysate (input), Lane 2: Rabbit anti-Cytokeratin 20 antibody IP in HT-29 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Cytokeratin 20 antibody in HT-29 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 3 seconds.| Product Name | Cytokeratin 20 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human Cytokeratin 20 aa 375-424 / 424. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:2000-1:5000 IHC:1:50-1:500 ICC/IF:1:50 FC:1:50 IP:1-2μg/sample |
|
| Species Reactivity | HumanRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | KRT20 |
|---|---|
| Gene Synonyms | K20 CD20 CK20 CK-20 KRT21 |
| Gene Full Name | keratin 20 |
| Gene Summary | The protein encoded by this gene is a member of the keratin family. The keratins are intermediate filament proteins responsible for the structural integrity of epithelial cells and are subdivided into cytokeratins and hair keratins. The type I cytokeratins consist of acidic proteins which are arranged in pairs of heterotypic keratin chains. This cytokeratin is a major cellular protein of mature enterocytes and goblet cells and is specifically expressed in the gastric and intestinal mucosa. The type I cytokeratin genes are clustered in a region of chromosome 17q12-q21. [provided by RefSeq, Jul 2008] |
| Molecular Weight(MW) | 48kDa(Observed band size: 48/50kDa) |
| Cellular Localization | Cytoplasm. |
WB
Western blot analysis of Cytokeratin 20 on different lysates with Rabbit anti-Cytokeratin 20 antibody at 1/5,000 dilution. Lane 1: HT-29 cell lysate, Lane 2: LoVo cell lysate, Lane 2: CRC cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 1 minute; 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 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit 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 Rabbit anti-Cytokeratin 20 antibody at 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 at 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.
IHC
Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-Cytokeratin 20 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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 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
ICC staining of Cytokeratin 20 in CRC 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 (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).
FC
Flow cytometric analysis of CRC cells with Cytokeratin 20 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
IP
Cytokeratin 20 was immunoprecipitated from 0.2 mg HT-29 cell lysate with Rabbit anti-Cytokeratin 20 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Cytokeratin 20 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HT-29 cell lysate (input), Lane 2: Rabbit anti-Cytokeratin 20 antibody IP in HT-29 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Cytokeratin 20 antibody in HT-29 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 3 seconds.| Application Notes | WB:1:2000-1:5000 IHC:1:50-1:500 ICC/IF:1:50 FC:1:50 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
