Recombinant Anti-RAB10 (phospho T73) antibody [MJF-R21] (ab230261)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MJF-R21] to RAB10 (phospho T73)
- Suitable for: WB, Dot blot
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-RAB10 (phospho T73) antibody [MJF-R21]
See all RAB10 primary antibodies -
Description
Rabbit monoclonal [MJF-R21] to RAB10 (phospho T73) -
Host species
Rabbit -
Tested applications
Suitable for: WB, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type MEF whole cell lysate; LRRK2 [R1441C] knock-in MEF whole cell lysate, 293T transfected with RAB10 expression vector containing a myc-His-tag®, whole cell lysate. Dot Blot: Rab10 (phospho T73) peptide.
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General notes
Please see PMID: 29127256. Lis P et al. Development of phospho-specific Rab protein antibodies to monitor in vivo activity of the LRRK2 Parkinson's disease kinase. Biochem J 475:1-22 (2018).
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This antibody was developed with support from The Michael J. Fox Foundation.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
MJF-R21 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab230261 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 |
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WB |
1/1000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
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Dot blot |
1/1000.
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Notes |
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WB
1/1000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa). |
Dot blot
1/1000. |
Target
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Function
The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different set of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion (By similarity). That Rab is mainly involved in the biosynthetic transport of proteins from the Golgi to the plasma membrane. Regulates, for instance, SLC2A4/GLUT4 glucose transporter-enriched vesicles delivery to the plasma membrane. In parallel, it regulates the transport of TLR4, a toll-like receptor to the plasma membrane and therefore may be important for innate immune response. Plays also a specific role in asymmetric protein transport to the plasma membrane within the polarized neuron and epithelial cells. In neurons, it is involved in axonogenesis through regulation of vesicular membrane trafficking toward the axonal plasma membrane while in epithelial cells, it regulates transport from the Golgi to the basolateral membrane. Moreover, may play a role in the basolateral recycling pathway and in phagosome maturation. According to PubMed:23263280, may play a role in endoplasmic reticulum dynamics and morphology controlling tubulation along microtubules and tubules fusion. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Cellular localization
Cytoplasmic vesicle membrane. Golgi apparatus membrane. Golgi apparatus, trans-Golgi network membrane. Endosome membrane. Recycling endosome membrane. Cytoplasmic vesicle, phagosome membrane. Cell projection, cilium. Endoplasmic reticulum membrane. Associates with SLC2A4/GLUT4 storage vesicles (PubMed:22908308). Localizes to the base of the cilium (PubMed:20576682). Transiently associates with phagosomes (By similarity). Localizes to the endoplasmic reticulum at domains of new tubule growth (PubMed:23263280). - Information by UniProt
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Database links
- Entrez Gene: 10890 Human
- Entrez Gene: 19325 Mouse
- Omim: 612672 Human
- SwissProt: P61026 Human
- SwissProt: P61027 Mouse
- Unigene: 467960 Human
- Unigene: 378993 Mouse
- Unigene: 486858 Mouse
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Alternative names
- GTP binding protein RAB10 antibody
- Rab10 antibody
- RAB10 member RAS oncogene family antibody
see all
Images
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All lanes : Anti-RAB10 (phospho T73) antibody [MJF-R21] (ab230261) at 1/1000 dilution
Lane 1 : 293T (Human embryonic kidney epithelial cell) transfected with an empty vector, containing a myc-His-tag®, whole cell lysate
Lane 2 : 293T transfected with RAB10 expression vector containing a myc-His-tag®, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 23 kDa
Exposure time: 180 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
Observed MW 28 kDa
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All lanes : Anti-RAB10 (phospho T73) antibody [MJF-R21] (ab230261) at 1/1000 dilution
Lane 1 : Wild-type MEF (mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : Wild-type MEF treated with 100 nM MLi-2 for 90 minutes, whole cell lysate
Lane 3 : LRRK2 [R1441C] knock-in MEF whole cell lysate
Lane 4 : LRRK2 [R1441C] knock-in MEF treated with 100 nM MLi-2 for 90 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 23 kDa
Observed band size: 23 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The LRRK2 pathogenic mutation R1441C increases LRRK2 activity and markedly elevates Rab10 phosphorylation in MEF (mouse embryonic fibroblasts).
The expression pattern is consistent with the literature (PMID: 29127256).
The cell lysates were kindly provided by our collaborator, Dr. Dario Alessi.
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All lanes : Anti-RAB10 (phospho T73) antibody [MJF-R21] (ab230261) at 1/1000 dilution
Lane 1 : Wild-type A549 (human lung carcinoma cell line) whole cell lysate
Lane 2 : Wild-type A549 treated with 100 nM MLi-2 for 90 minutes, whole cell lysate
Lane 3 : Rab8A knock-out A549 whole cell lysate
Lane 4 : Rab8A knock-out A549 treated with 100 nM MLi-2 for 90 minutes, whole cell lysate
Lane 5 : Rab10 knock-out A549 whole cell lysate
Lane 6 : Rab10 knock-out A549 treated with 100 nM MLi-2 for 90 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-labeled secondary antibody at 1/2500 dilution
Predicted band size: 23 kDa
Observed band size: 23 kDaBlocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% BSA/TBST.
The images were kindly provided by our collaborator, Dr. Dario Alessi, and have been published (PMID: 29127256).
Scanned with Licor Odyssey CLx.
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All lanes : Anti-RAB10 (phospho T73) antibody [MJF-R21] (ab230261) at 1/1000 dilution
Lane 1 : HEK-293 (human epithelial cell line from embryonic kidney) cells transfected with LRRK2 [Y1699C] and HA-tagged Rab3A expression vectors, were treated with 150 nM MLi-2 for 90 minutes, whole cell lysate
Lane 2 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab8A expression vectors, were treated with 150 nM MLi-2 for 90 minutes, whole cell lysate
Lane 3 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab10 expression vectors, were treated with 150 nM MLi-2 for 90 minutes, whole cell lysate
Lane 4 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab35 expression vectors, were treated with 150 nM MLi-2 for 90 minutes, whole cell lysate
Lane 5 : HEK-293 cells transfected with LRRK2[Y1699C] and HA-tagged Rab43 expression vectors, were treated with 150 nM MLi-2 for 90 minutes, whole cell lysate
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : IRDye 800CW secondary antibody at 1/25000 dilution
Predicted band size: 23 kDa
Observed band size: 23 kDaBlocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% BSA/TBST.
The LRRK2 pathogenic mutation Y1699C increases LRRK2 activity and markedly elevates the phosphorylation of Rab proteins.
The images were kindly provided by our collaborator Dr. Dario Alessi, and have been published (PMID: 29127256).
Scanned with Licor Odyssey CLx.
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Dot blot analysis of Rab10 (phospho T73) labeled with ab230261 at 1/1000 dilution.
Lane 1: Rab10 (phospho T73) peptide;
Lane 2: Rab10 non-phospho peptide.
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100,000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 32 seconds.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (50)
ab230261 has been referenced in 50 publications.
- Liu Z et al. α-Synuclein-containing erythrocytic extracellular vesicles: essential contributors to hyperactivation of monocytes in Parkinson's disease. J Neuroinflammation 19:53 (2022). PubMed: 35193594
- Kumar R et al. A cell-based GEF assay reveals new substrates for DENN domains and a role for DENND2B in primary ciliogenesis. Sci Adv 8:eabk3088 (2022). PubMed: 35196081
- Singh RK et al. Nanobodies as allosteric modulators of Parkinson's disease-associated LRRK2. Proc Natl Acad Sci U S A 119:N/A (2022). PubMed: 35217606
- Stormo AED et al. The E3 ligase TRIM1 ubiquitinates LRRK2 and controls its localization, degradation, and toxicity. J Cell Biol 221:N/A (2022). PubMed: 35266954
- Fernández B et al. Evaluation of Current Methods to Detect Cellular Leucine-Rich Repeat Kinase 2 (LRRK2) Kinase Activity. J Parkinsons Dis 12:1423-1447 (2022). PubMed: 35599495