Immunofluorescence analysis of RYK was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RYK (F35 P7 D7 F5) Mouse Monoclonal Antibody (MA1615) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Clone||F35 P7 D7 F5|
|Immunogen||Synthetic peptide corresponding to residues C(462) L T E F H A A L G A Y V(475) of RYK, conjugated to ovalbumin.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||2 ug/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody detects RYK in human, mouse, and rat samples.
MA1-615 has been successfully used in Western blot, procedures. By Western blot this antibody detects the RYK receptor-like tyrosine kinase-1 in cells.
The MA1-615 immunogen corresponds to the ovalbumin-conjugated synthetic peptide C (462) L T E F H A A L G A Y V(475).
RYK (Receptor-like Tyrosine kinase-1) is in the Tyrosine family of receptor kinases. It is found in the plasma membrane of heart, prostate and kidney cells. RYK contains 604 amino acids and has an approximate molecular weight of 67507 Da. It contains one WIF domain. It functions as a potential growth factor receptor protein tyrosine kinase. RYK are divergent from those of other classic receptor tyrosine kinases. It is an orphan receptor that contains a catalytically inactive tyrosine-kinase-related domain. RYK is expressed in nestin-positive progenitor cells and neurons, and in a certain population of oligodendrocytes, O-2A progenitor cells, and type-2 astrocytes in developing CNS.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.