Name: Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)]
Brand: RabMAb
SKU: ab133450

Product name

Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)]
See all LRRK2 primary antibodies
Description

Rabbit monoclonal [UDD2 10(12)] to LRRK2 (phospho S935)
Host species

Rabbit
Specificity

The antibody does not give a positive signal in U-87 MG, SH-SY-5Y and human fetal brain. Please contact our Scientific Support team if you have any question.
Tested applications

Suitable for: WB, ICC/IFmore details
Unsuitable for: IHC-P
Species reactivity

Reacts with: Mouse, Human
Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human LRRK2 (phospho S935).
Database link: Q5S007
Positive control
- WB: GFP LRRK2, GFP LRRK2 S910A, GFP LRRK2 S935A, LRRK2 WT MEF, LRRK2 WT MEF, Lymphoblastoid and RAW 264.7 cell lysates. ICC/IF: A431 cells.
General notes

This antibody was developed with the support of The Michael J. Fox Foundation (MJFF) and in partnership with Dr. Dario Alessi (MRC Protein Phosphorylation Unit, University of Dundee) to help accelerate LRRK2 research. Dr. Alessi has characterized several unique and high quality LRRK2 rabbit monoclonal antibodies, generated by Epitomics, to be made widely available for PD research community.

LRRK2 (Leucine-rich repeat kinase 2, dardarin) is a multi-domain protein belonging to the ROCO family of proteins that contains a kinase and GTPase domain among its many protein interaction domains. LRRK2 is mutated in a significant number of Parkinson's disease (PD) patients. Mutations in this gene account for 4% of PD, and are observed in 1% of sporadic PD patients. The most common mutation replaces glycine 2019 with a serine that results in increased LRRK2 kinase activity. This indicates that inhibitors of LRRK2 kinase activity might be of therapeutic benefit for the treatment of Parkinson’s disease and has stimulated much activity in this field of research.
Recent work has revealed that LRRK2 interacts with 14-3-3 phospho-binding adaptor isoforms that is mediated by phosphorylation of Ser910 and Ser935 located prior to the leucine rich repeat domain mediates. Interestingly, 14-3-3 binding has been linked to Parkinson’s disease as Ser910 as well as Ser935 and interaction with the 14-3-3 is inhibited by five of the six validated LRRK2 pathogenic mutations (R1441C, R1441G, R1441H, Y1699C and I2020T).

The Dundee-MJFF LRRK2 PhosphoSer935 antibody will be of great utility in further understanding the link between 14-3-3 binding to LRRK2 and Parkinson’s disease as well as assessing the efficacy of LRRK2 inhibitors that are being developed. It should be noted the Dundee-MJFF antibody is highly selective and sensitive and can readily be used to monitor LRRK2 Ser935 phosphorylation in immunoblot analysis of 2-20 microgram amounts of whole cell extract. The Dundee-MJFF LRRK2 PhosphoSer935 recognizes both mouse and human endogenous LRRK2.

A BSA and Azide free version of this product is available as ab172382.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor^® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

UDD2 10(12)
Isotype

IgG
Research areas

Alternative Versions
- Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] - BSA and Azide free (ab172382)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Immunizing Peptide (Blocking)
- Human LRRK2 (phospho S935) peptide (ab229694)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human LRRK2 protein (ab169908)

Our Abpromise guarantee covers the use of ab133450 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Detects a band of approximately 286 kDa (predicted molecular weight: 286 kDa).Can be blocked with Human LRRK2 (phospho S935) peptide (ab229694).
ICC/IF		1/100.

Application notes

Is unsuitable for IHC-P.

Function

Positively regulates autophagy through a calcium-dependent activation of the CaMKK/AMPK signaling pathway. The process involves activation of nicotinic acid adenine dinucleotide phosphate (NAADP) receptors, increase in lysosomal pH, and calcium release from lysosomes. Together with RAB29, plays a role in the retrograde trafficking pathway for recycling proteins, such as mannose 6 phosphate receptor (M6PR), between lysosomes and the Golgi apparatus in a retromer-dependent manner. Regulates neuronal process morphology in the intact central nervous system (CNS). Plays a role in synaptic vesicle trafficking. Phosphorylates PRDX3. Has GTPase activity. May play a role in the phosphorylation of proteins central to Parkinson disease.
Tissue specificity

Expressed in the brain. Expressed in pyramidal neurons in all cortical laminae of the visual cortex, in neurons of the substantia nigra pars compacta and caudate putamen (at protein level). Expressed throughout the adult brain, but at a lower level than in heart and liver. Also expressed in placenta, lung, skeletal muscle, kidney and pancreas. In the brain, expressed in the cerebellum, cerebral cortex, medulla, spinal cord occipital pole, frontal lobe, temporal lobe and putamen. Expression is particularly high in brain dopaminoceptive areas.
Involvement in disease

Parkinson disease 8
Sequence similarities

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
Contains 12 LRR (leucine-rich) repeats.
Contains 1 protein kinase domain.
Contains 1 Roc domain.
Contains 7 WD repeats.
Domain

The seven-bladed WD repeat region is critical for synaptic vesicle trafficking and mediates interaction with multiple vesicle-associated presynaptic proteins.
The Roc domain mediates homodimerization and regulates kinase activity.
Post-translational
modifications

Autophosphorylated.
Cellular localization

Membrane. Cytoplasm. Perikaryon. Mitochondrion. Golgi apparatus. Cell projection, axon. Cell projection, dendrite. Endoplasmic reticulum. Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. Endosome. Lysosome. Mitochondrion outer membrane. Mitochondrion inner membrane. Mitochondrion matrix. Predominantly associated with intracytoplasmic vesicular and membranous structures (By similarity). Localized in the cytoplasm and associated with cellular membrane structures. Predominantly associated with the mitochondrial outer membrane of the mitochondria. Colocalized with RAB29 along tubular structures emerging from Golgi apparatus. Localizes in intracytoplasmic punctate structures of neuronal perikarya and dendritic and axonal processes.
Target information above from: UniProt accession Q5S007 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 120892 Human
- Entrez Gene: 66725 Mouse
- Omim: 609007 Human
- SwissProt: Q5S007 Human
- SwissProt: Q5S006 Mouse
- Unigene: 187636 Human
- Unigene: 37558 Mouse
Alternative names
- augmented in rheumatoid arthritis 17 antibody
- AURA17 antibody
- Dardarin antibody
- Leucine rich repeat kinase 2 antibody
- leucine rich repeat serine threonine protein kinase 2 antibody
- Leucine-rich repeat serine/threonine-protein kinase 2 antibody
- LRRK 2 antibody
- LRRK2 antibody
- LRRK2_HUMAN antibody
- PARK 8 antibody
- PARK8 antibody
- RIPK7 antibody
- ROCO 2 antibody
- ROCO2 antibody
see all

Western blot - Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450)Ito et al PLoS One. 2014 May 16;9(5):e97988. doi: 10.1371/journal.pone.0097988. eCollection 2014. Fig 3.

Inhibitor-induced dephosphorylation of kinase-inactive LRRK2.

HEK-293 cells transfected with wild-type, K1906A, K1906M, D1994A, D1994N, D2017A, S2032A or T2035A LRRK2 were treated with (Panel A) 3 µM LRRK2-IN-1 or the solvent (0.1% DMSO) for 30 min.
Immunocytochemistry/ Immunofluorescence - Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450)

Immunocytochemistry/Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labeling LRRK2 (phospho S935) with purified ab133450 at 1/100.

Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control: Secondary antibody, ab150113, an Alexa Fluor^® 488-conjugated goat anti-mouse IgG (1/500).
Western blot - Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450)

All lanes : Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450) at 1/1000 dilution (unpurified)

Lane 1 : GFP LRRK2 lysate at 5 µg
Lane 2 : GFP LRRK2 S910A lysate at 5 µg
Lane 3 : GFP LRRK2 S935A lysate at 5 µg
Lane 4 : LRRK2 WT MEF lysate at 20 µg
Lane 5 : LRRK2 WT MEF lysate from LRRK2 IN1 treated cells
at 20 µg
Lane 6 : LRRK2 KO MEF lysate
at 20 µg
Lane 7 : LRRK2 KO MEF lysate from LRRK2 IN1 treated cells
at 20 µg
Lane 8 : Lymphoblastoid lysate
at 30 µg
Lane 9 : Lymphoblastoid lysate from LRRK2 IN1 treated cells
at 30 µg

Secondary
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Predicted band size: 286 kDa
Western blot - Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450)

All lanes : Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450) at 1/5000 dilution (purified)

Lane 1 : WT-LRRK2 cell lysate - untreated
Lane 2 : WT-LRRK2 cell lysate - treated with Lambda phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 286 kDa
Observed band size: 286 kDa

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Western blot - Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450)

Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450) at 1/1000 dilution (purified) + RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 286 kDa
Observed band size: 286 kDa

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

Western blot protocols
Immunocytochemistry & immunofluorescence protocols

This product has been referenced in:

Henderson MX et al. LRRK2 inhibition does not impart protection from a-synuclein pathology and neuron death in non-transgenic mice. Acta Neuropathol Commun 7:28 (2019). Read more (PubMed: 30808409) »
Bae EJ et al. LRRK2 kinase regulates a-synuclein propagation via RAB35 phosphorylation. Nat Commun 9:3465 (2018). Read more (PubMed: 30150626) »

See all 31 Publications for this product

Publishing research using ab133450? Please let us know so that we can cite the reference in this datasheet.

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Recombinant Anti-LRRK2 (phospho S935) antibody [UDD2 10(12)] (ab133450)

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