Immunofluorescence analysis of LRRK2 [pS935] was done on 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Phospho-LRRK2 / PARK8 pSer935 (3H8L19), ABfinity™ Rabbit Monoclonal Antibody (701066) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphopeptide corresponding to amino acids 930–940 of human LRRK2|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||2-5 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 is predicted to react with non-human primate, mouse and rat based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
LRRK2 belongs to the leucine-rich repeat kinase family commonly known as Dardarin. The LRRK2 gene encodes for a protein with an ankyrin-repeat region, a leucine-rich repeat region, a kinase domain, a RAS domain, a GTPase-domain, a MLK-like domain, and a WD40 domain. Proteins with leucine-rich regions play important roles in activities that require protein-protein interactions, such as transmitting signals or helping to assemble the cell's structural framework. Phosphorylation of LRRK2 at multiple sites including serine 910 and serine 935 regulates LRRK2 mediated cellular activities by affecting binding of dimeric 14-3-3 protein. Phosphorylation at Ser910, found to be constitutive in various tissues, is caused by protein kinase A. In humans, he gene is present on the q arm of chromosome 12.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.