Antibodies, Proteins, Kits and Reagents for Life Science

154 118 ₸ Product size

100 µl 154 118 ₸

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Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [MJF-R20] to RAB8A (phospho T72)
Suitable for: WB, Dot blot
Reacts with: Mouse, Rat, Human

Product name

Anti-RAB8A (phospho T72) antibody [MJF-R20]
See all RAB8A primary antibodies
Description

Rabbit monoclonal [MJF-R20] to RAB8A (phospho T72)
Host species

Rabbit
Specificity

This 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 Applications & Species

Application Species
Dot
Rat

WB
Mouse

See all applications and species data
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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.

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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Application	Species
Dot	Rat
WB	Mouse

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 (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

MJF-R20
Isotype

IgG
Research areas

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 kDa

Blocking 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)

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 minute

Blocking/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)

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.
Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)

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