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Synthetic peptide, corresponding to residues in Human RYK (intracellular domain).
This product is a recombinant rabbit monoclonal antibody.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
Our Abpromise guarantee covers the use of ab124961 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/50000 - 1/200000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).|
|IP||1/10 - 1/100.|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes)|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/100 - 1/250.|
Purified ab124961 staining RYK in Jurkat (Human T cell leukemia T lymphocyte) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody using purified anti-RYK, ab124961. An AlexaFluor®488 Goat anti-Rabbit was used as the secondary antibody. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain antibody. DAPI was used as a nuclear counter stain. Confocal image showing nuclear and cytoplasmic staining in Jurkat cells.
Purified ab124961 staining RYK in Human ovarian carcinoma tissue sections by Immunohistochemistry (IHC-P- paraformaldehye- fixed, paraffin embedded sections). Tissue was fixed with paraffin and heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antbody at 1:400 dilution. A ready to use Goat Anti- Rabbit H&L (HRP). Hematoxylin was used as a counterstain. Cytoplasmic staining on human ovarian carcinoma.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure: Lane 1: 30 seconds
Lane 2: 10 seconds
Purified ab124961 staining RYK in Jurkat (Human T cell leukemia T lymphocyte) cell line by Flow Cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Samples were incubated with primary antibody at 1:40 dilution (red). An Alexa Fluor® 488 Goat anti-rabbit IgG , ab150077 was used as a secondary antibody at 1:2000 dilution. A rabbit monoclonal IgG, ab172730 was used as an isotype control (black). Cell without incubation with primary and secondary antibodies - unlabeled control (blue).
Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate, 10μg
Lane 2 (+): HEK-293 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124961 in HEK-293 whole cell lysate
Purified ab124961 immunoprecipitating in HEK-293 whole cell lysate. For western blotting, primary antibody used was purified ab124961 at 1:500 dilution. Ab131366 VeriBlot for IP (HRP) was used as the secondary antibody at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST