Cytokeratin 18 KRT18 Antibody(monoclonal, 7I6)

• Catalog Number: orb623836
• Category: Antibodies
• Description: Cytokeratin 18 KRT18 Antibody(monoclonal, 7I6)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 7I6
• Tested applications: FC, ICC, IF, IHC, IHC-Fr, WB
• Reactivity: Human
• Isotype: Mouse IgG1
• Immunogen: E.coli-derived human Cytokeratin 18 recombinant protein (Position: E204-H430). Human Cytokeratin 18 shares 87.7% and 85.9% amino acid (aa) sequence identity with mouse and rat Cytokeratin 18, respectively.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.5μg/ml, Human Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Immunofluorescence, 2μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 48 kDa
• UniProt ID: P05783
• Sensitivity: > 5000 cells
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Alternative names: Keratin, type I cytoskeletal 18; Cell proliferatio
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. . Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
• Expiration Date: 12 months from date of receipt.

IHC analysis of Cytokeratin 18 using anti-Cytokeratin 18 antibody (orb623836). Cytokeratin 18 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cytokeratin 18 Antibody (orb623836) 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) (Catalog # orb90443) with DAB as the chromogen.

IHC analysis of Cytokeratin 18 using anti-Cytokeratin 18 antibody (orb623836). Cytokeratin 18 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cytokeratin 18 Antibody (orb623836) 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) (Catalog # orb90443) with DAB as the chromogen.

IF analysis of Cytokeratin 18 using anti-Cytokeratin 18 antibody (orb623836). Cytokeratin 18 was detected in immunocytochemical section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-Cytokeratin 18 Antibody (orb623836) overnight at 4°C. DyLight®488 Conjugated Goat Anti-mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of Cytokeratin 18 using anti-Cytokeratin 18 antibody (orb623836). Cytokeratin 18 was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL mouse anti-Cytokeratin 18 Antibody (orb623836) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Cytokeratin 18 using anti-Cytokeratin 18 antibody (orb623836). 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 50ug of sample under reducing conditions. Lane 1: human placenta tissue lysates, Lane 2: human Caco-2 whole cell lysates, Lane 3: human A549 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 hours at RT. The membrane was incubated with mouse anti-Cytokeratin 18 antigen affinity purified monoclonal antibody (Catalog # orb623836) 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for Cytokeratin 18 at approximately 48KD. The expected band size for Cytokeratin 18 is at 48KD.

Flow Cytometry analysis of A431 cells using anti-Cytokeratin 18 antibody (orb623836). Overlay histogram showing A431 cells stained with orb623836 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytokeratin 18 Antibody (orb623836, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.