The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 - 10 µg/ml.
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
Use at an assay dependent concentration.
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 5 - 10 µg/ml.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Involved in the organization of myofibers. Together with KRT8, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.
Expressed in a defined zone of basal keratinocytes in the deep outer root sheath of hair follicles. Also observed in sweat gland and mammary gland ductal and secretory cells, bile ducts, gastrointestinal tract, bladder urothelium, oral epithelia, esophagus, ectocervical epithelium (at protein level). Expressed in epidermal basal cells, in nipple epidermis and a defined region of the hair follicle. Also seen in a subset of vascular wall cells in both the veins and artery of human umbilical cord, and in umbilical cord vascular smooth muscle. Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma in structures that contain dystrophin and spectrin.
Belongs to the intermediate filament family.
Present in hair follicles at all stages of development.
This keratin differs from all other IF proteins in lacking the C-terminal tail domain.
Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 19 antibody [BA16] (ab20210)This image is a courtesy of Rene Villadsen
ab20210 staining Cytokeratin 19 in human breast carcinoma tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with methanol and blocking with 10% serum for 5 minutes was performed. The sample was incubated with primary antibody (1/100) in PBS with 10% goat serum for 1 hour. A HRP-conjugated rabbit polyclonal to mouse IgG was used undiluted as secondary antibody.
Immunocytochemistry - Anti-Cytokeratin 19 antibody [BA16] (ab20210)This image is courtesy of an Abreview submitted by Rene Villadsen
ab20210 at 1/100 dilution staining Cytokeratin 19 breast cancer cell line by immunocytochemistry. Sections were methanol fixed prior to blocking in 10% serum for 5 minutes and then incubated with ab20210, for 1 hour at 25°C. Ultravision ONE HRP rabbit polyclonal to mouse Ig was used as the secondary antibody.
ab20210 stained HepG2 cells. The cells were 100% methanol fixed for 5 minutes at room temperature and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab20210 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 used at a 1/1000 dilution for 1hour at room temperature. The cells were counterstained with ab202277 (Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 594) (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
IHC image of Cytokeratin 19 staining in human normal liver formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20210, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Cytokeratin 19 antibody [BA16] (ab20210)
All lanes : Anti-Cytokeratin 19 antibody [BA16] (ab20210) at 1 µg/ml
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 2 : Human liver tissue lysate - total protein (ab29889) Lane 3 : Human tonsil normal tissue lysate - total protein (ab29615)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution