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- Western blot [3]
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- Product number
- PA5-22125 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cytokeratin 20 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLa.
- Concentration
- 0.88 mg/mL
Submitted references The mitochondrial deoxyguanosine kinase is required for cancer cell stemness in lung adenocarcinoma.
Lin S, Huang C, Sun J, Bollt O, Wang X, Martine E, Kang J, Taylor MD, Fang B, Singh PK, Koomen J, Hao J, Yang S
EMBO molecular medicine 2019 Dec;11(12):e10849
EMBO molecular medicine 2019 Dec;11(12):e10849
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Cytokeratin 20 Polyclonal Antibody (Product # PA5-22125). Sample (30 µg of whole cell lysate). Lane A: HeLa. 10% SDS PAGE. Cytokeratin 20 Polyclonal Antibody (Product # PA5-22125) diluted at 1:3,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of KRT20 was achieved by transfecting HT 29 cells with KRT20 specific siRNAs (Silencer® select Product # s29040, Product # s29041). Western blot analysis (Fig. a) was performed using whole cell extracts from the KRT20 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed using KRT20 Polyclonal Antibody (Product # PA5-22125) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to KRT20.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Cytokeratin 20 Polyclonal Antibody (Product # PA5-22125) on whole cell extracts (30 µg lysate) of HT-29 (Lane 1), HCT-116 (Lane 2) and MCF7 (Lane 3) and 48.48 kDa band corresponding to Cytokeratin 20 was observed only in HT-29 which is reported to be the positive model. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Cytokeratin 20 in paraformaldehyde-fixed HeLa cells using a Cytokeratin 20 polyclonal antibody (Product # PA5-22125) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using Cytokeratin 20 (Product # PA5-22125) antibody at 1:200 dilution.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Cytokeratin 20 Polyclonal Antibody detects Cytokeratin 20 protein at cytoplasm on mouse urinary bladder by immunohistochemical analysis. Sample: Paraffin-embedded mouse urinary bladder. Cytokeratin 20 Polyclonal Antibody (Product # PA5-22125) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded HT29 xenograft, using Cytokeratin 20 (Product # PA5-22125) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 DGUOK is required for cancer cell stemness in lung adenocarcinoma cells A, B Flow cytometric assay using ALDEFLUOR (A) and anti-CD166 antibody (B) showing the effects of DGUOK depletion on the proportion of ALDH + cells (A) and CD166 + cells (B) in lung adenocarcinoma cell lines. C, D Representative images and quantitation of the effects of DGUOK depletion on lung tumor sphere formation by lung adenocarcinoma cells. E Cell survival assay using annexin V/PI staining and flow cytometric to determine the effects of DGUOK depletion on H1650 cell survival under adherent and non-adherent (30 h in lung sphere formation conditions) conditions. F Quantitation of apoptotic cells in (E) from three independent biological replicates. G, H Representative immunofluorescence staining showing the effects of DGUOK KO on expression of CK20 in Matrigel-embedded H1650 (G) and A549 (H) tumor spheres. FBS was added at day 5 to induce CD20 expression. Data information: Data are shown as mean +- SD. n = 3 in (D and F). P values were determined by two-tailed, two-sample Student's t -test. Experiments were repeated with three biological replicates with similar results.
