Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-27250 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cytokeratin 20 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Recommended positive controls: HeLaS3. Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Cytokeratin 20 Polyclonal Antibody (Product # PA5-27250). Sample (30 µg whole cell lysate). A: HeLa S3. 10% SDS PAGE. Cytokeratin 20 Polyclonal Antibody (Product # PA5-27250) diluted at 1:1,000.
- 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 # PA5-27250) 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 # PA5-27250) 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
- Immunohistochemical analysis of paraffin-embedded HT29 xenograft , using Cytokeratin 20 (Product # PA5-27250) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.