Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)
- Datasheet
- References (1)
- Protocols
Overview
-
Product nameAnti-RAB8A (phospho T72) antibody [MJF-R20]
See all RAB8A primary antibodies -
DescriptionRabbit monoclonal [MJF-R20] to RAB8A (phospho T72)
-
Host speciesRabbit
-
SpecificityThis antibody cross-reacts with phosphorylated Rab3A, Rab10, Rab35 and Rab43. Please note that the immunogen sequence was derived from Rab8A but is identical to that of Rab8B.
-
Tested applicationsSuitable for: WB, Dot blotmore details
-
Species reactivityReacts with: Mouse, Human
-
Immunogen
Synthetic peptide within Human RAB8A aa 50-150. The exact sequence is proprietary. The immunogen sequence of Rab8A is identical to Rab8B.
Database link: P61006 -
Positive control
- Dot Blot: Rab8 (phospho T72) peptide. WB: LRRK2 [R1441C] knock-in MEF (immortalized), whole cell lysate. HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab8A, Rab10 and Rab35 expression vectors, treated with 150 nM MLi-2.
-
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 antibody was developed with support from The Michael J. Fox Foundation.
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 product is a recombinant rabbit monoclonal antibody.
Properties
-
FormLiquid
-
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
-
ClonalityMonoclonal
-
Clone numberMJF-R20
-
IsotypeIgG
-
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Related Products
Applications
Our Abpromise guarantee covers the use of ab230260 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. Predicted molecular weight: 24 kDa. | |
| Dot blot | 1/1000. |
Target
-
FunctionMay be involved in vesicular trafficking and neurotransmitter release. Together with RAB11A, RAB3IP, the exocyst complex, PARD3, PRKCI, ANXA2, CDC42 and DNMBP promotes transcytosis of PODXL to the apical membrane initiation sites (AMIS), apical surface formation and lumenogenesis. Together with MYO5B and RAB11A participates in epithelial cell polarization.
-
Sequence similaritiesBelongs to the small GTPase superfamily. Rab family.
-
Cellular localizationCell membrane. Golgi apparatus. Cytoplasm > perinuclear region. Cell projection. Colocalizes with OPTN at the Golgi complex and in vesicular structures close to the plasma membrane. In the GDP-bound form, present in the perinuclear region. Shows a polarized distribution to distal regions of cell protrusions in the GTP-bound form. Colocalizes with PARD3, PRKCI, EXOC5, OCLN, PODXL and RAB11A in apical membrane initiation sites (AMIS) during the generation of apical surface and luminogenesis.
- Information by UniProt
-
Database links
- Entrez Gene: 4218 Human
- Entrez Gene: 17274 Mouse
- Omim: 165040 Human
- SwissProt: P61006 Human
- SwissProt: P55258 Mouse
- Unigene: 642874 Human
- Unigene: 709846 Human
- Unigene: 162811 Mouse
-
Alternative names
- AA409338 antibody
- MEL antibody
- Mel transforming oncogene (derived from cell line NK14) RAB8 homolog antibody
see all
Images
-
![Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)](https://www.abcam.com/ps/products/230/ab230260/Images/ab230260-315217-wbupdatedlaneimage.jpg)
Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)All lanes : Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260) at 1/1000 dilution
Lane 1 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab3A expression vectors, were treated with150 nM MLi-2 for 90 minutes, whole cell lysate, 100 ng
Lane 2 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab8A expression vectors, were treated with150 nM MLi-2 for 90 minutes, whole cell lysate, 100 ng
Lane 3 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab10 expression vectors, were treated with150 nM MLi-2 for 90 minutes, whole cell lysate, 100 ng
Lane 4 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab35 expression vectors, were treated with150 nM MLi-2 for 90 minutes, whole cell lysate, 100 ng
Lane 5 : HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab43 expression vectors, were treated with150 nM MLi-2 for 90 minutes, whole cell lysate, 100 ng
Secondary
All lanes : IRDye 800CW secondary antibody at 1/25000 dilution
Predicted band size: 24 kDaBlocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% BSA/TBST.
The LRRK2 pathogenic mutation Y1699C increases LRRK2 activity and markedly elevates Rab8 phosphorylation in MEF (mouse embryonic fibroblasts).
The results show that this antibody recognizes other LRRK2-phosphorylated Rab proteins (Rab3A, Rab10, and Rab35). The phospho-threonine site in Rab8 (T72) differs from that of Rab3A (T86); Rab10 (T73); and Rab43 (T82).
The images were kindly provided by our collaborator, Dr. Dario Alessi, and have been published (PMID: 29127256).
-
![Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)](https://www.abcam.com/ps/products/230/ab230260/Images/ab230260-2-230260WB1.jpg)
Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)All lanes : Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260) at 1/1000 dilution
Lane 1 : Wild-type MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate
Lane 2 : Wild-type MEF (mouse embryonic fibroblast (immortalized)) treated with 100 nM MLi-2 for 90 minutes, whole cell lysate
Lane 3 : LRRK2 [R1441C] knock-in MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate
Lane 4 : LRRK2 [R1441C] knock-in MEF (mouse embryonic fibroblast (immortalized)) 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
Predicted band size: 24 kDa
Exposure time: 1 minuteBlocking/Dilution buffer and concentration: 5% NFDM/TBST.
The LRRK2 pathogenic mutation R1441C increases LRRK2 activity and markedly elevates Rab8 phosphorylation in MEF (mouse embryonic fibroblasts).
The expression profile is consistent with the literature (PMID: 29127256).
The cell lysates were kindly provided by our collaborator, Dr. Dario Alessi.
-
![Dot Blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)](https://www.abcam.com/ps/products/230/ab230260/Images/ab230260-1-230260Dot1.jpg)
Dot Blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)Dot Blot - Anti-Rab8A (phospho T72) antibody [MJF-R20] (ab230260) used at a 1/1000 dilution.
Lane 1: Rab8 (phospho T72) peptide.
Lane 2: Rab8 non-phospho peptide.
ab97051 secondary antibody used at a 1/100,000 dilution.
Exposure time: 32 seconds.
Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Datasheets and documents
References
This product has been referenced in:
- Lis P et al. Development of phospho-specific Rab protein antibodies to monitorin vivoactivity of the LRRK2 Parkinson's disease kinase. Biochem J 475:1-22 (2018). Read more (PubMed: 29127256) »
