Anti-Cytokeratin 19 KRT19 Antibody (monoclonal, 3D4)

• Catalog Number: orb421122
• Category: Antibodies
• Description: Anti-Cytokeratin 19 KRT19 Antibody (monoclonal, 3D4). Tested in IF, IHC, WB applications. This antibody reacts with Human.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 3D4
• Tested applications: IF, IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG1
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Cytokeratin 19, different from the related mouse and rat sequences by nine amino acids.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 44 kDa
• UniProt ID: P08727
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Keratin, type I cytoskeletal 19; Cytokeratin-19; C
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Immunofluorescence, 2μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of Cytokeratin 19 using anti-Cytokeratin 19 antibody. Cytokeratin 19 was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/mL mouse anti-Cytokeratin 19 Antibody overnight at 4°C. DyLight®550 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:200 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Cytokeratin 19 using anti-Cytokeratin 19 antibody. Cytokeratin 19 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Cytokeratin 19 Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of Cytokeratin 19 using anti-Cytokeratin 19 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human placenta tissue lysates, Lane 3: human SK-OV-3 whole cell lysates, Lane 4: human COLO-320 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Cytokeratin 19 antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system.