PA5-32463

antibody from Invitrogen Antibodies

Targeting: KRT19 CK19, K19, K1CS, MGC15366

Antibody data

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Validation data

Product number: PA5-32463 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cytokeratin 19 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is skin tissue.
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 500 µL
Concentration: 0.04 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

KRT19 protein structure - PA5-32463 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of KRT19 was achieved by transfecting MCF-7 with KRT19 specific siRNAs (Silencer® select Product # s7998). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the KRT19 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cytokeratin 19 Polyclonal Antibody (Product # PA5-32463, 1 ug/ml) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 KRT19.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Cytokeratin 19 Polyclonal Antibody (Product # PA5-32463) and a 44 kDa band corresponding to KRT19 was observed across cell lines tested. An additional band around 38 kDa has also been observed which is reported for Cytokeratin 19. Membrane enriched extracts (30 µg lysate) of MCF-7 (Lane 1), SK-BR-3 (Lane 2), T-47D (Lane 3), HT-29 (Lane 4), COLO 205 (Lane 5) and U-2 OS (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 ug/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (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).