AB231706

Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free

RabMAb
Recombinant
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(4 Publications)

Rabbit Recombinant Monoclonal RAB8A phospho T72 antibody. Carrier free. Suitable for Dot, WB and reacts with Synthetic peptide, Mouse, Human, Rat samples. Cited in 4 publications.

View Alternative Names

MEL, RAB8, RAB8A, Ras-related protein Rab-8A, Oncogene c-mel

2 Images

PubMed

Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free (AB231706)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230260).

All lanes:

Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free (ab231706) 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

false

Unknown

Dot Blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free (AB231706)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230260).

Unconjugated

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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

MJF-R20

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

Dot, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

ab230260 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.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Rat": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "guaranteed", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Synthetic peptide": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }

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Product details

ab231706 is the carrier-free version of ab230260.

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).

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

RAB8A is a small GTPase also referred to as Ras-related protein Rab-8A involved in intracellular membrane trafficking. It plays a critical role in the regulation of exocytosis the process by which cells release substances to the extracellular space. RAB8A mainly functions by cycling between an inactive GDP-bound form and an active GTP-bound form facilitating vesicle budding transport and fusion with target membranes. This protein is expressed in various tissues including the brain and pancreas where it assists in important cellular functions. RAB8A has a molecular mass of approximately 24 kDa.

Biological function summary

The small GTPase RAB8A contributes to the biosynthetic transport from the trans-Golgi network to the plasma membrane. It performs this activity in coordination with other proteins as part of a larger complex. RAB8A also plays an essential role in cilia formation and maintenance affecting cell polarity and signaling functions. The protein's ability to regulate vesicle movement makes it important for cellular homeostasis and communication.

Pathways

The regulatory function of RAB8A is significant in the ciliary membrane trafficking pathway and the insulin signaling pathway. Within these pathways RAB8A interacts closely with other proteins like Rab11 and Rabin8 which help support its roles in directing vesicle transport. These interactions are essential for proper cellular signaling and the maintenance of polarized cellular structures further supporting the function of various cellular pathways.

The dysfunction of RAB8A has associations with certain ciliopathies notably Bardet-Biedl syndrome. This condition involves abnormalities in cilia structure and function leading to symptoms like kidney disease vision impairment and obesity. Moreover disruptions in RAB8A have connections with insulin resistance implicating it in metabolic disorders. In the context of these diseases proteins such as BBS proteins and Rab11 play a role in mediating the effects of dysfunctional RAB8A-related pathways.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

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 sets of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion. RAB8A is involved in polarized 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 (PubMed : 20890297). Regulates the compacted morphology of the Golgi (PubMed : 26209634). Together with MYO5B and RAB11A participates in epithelial cell polarization (PubMed : 21282656). Also involved in membrane trafficking to the cilium and ciliogenesis (PubMed : 21844891, PubMed : 30398148, PubMed : 20631154). Together with MICALL2, may also regulate adherens junction assembly (By similarity). May play a role in insulin-induced transport to the plasma membrane of the glucose transporter GLUT4 and therefore play a role in glucose homeostasis (By similarity). Involved in autophagy (PubMed : 27103069). Participates in the export of a subset of neosynthesized proteins through a Rab8-Rab10-Rab11-dependent endososomal export route (PubMed : 32344433). Targeted to and stabilized on stressed lysosomes through LRRK2 phosphorylation (PubMed : 30209220). Suppresses stress-induced lysosomal enlargement through EHBP1 and EHNP1L1 effector proteins (PubMed : 30209220).

See full target information RAB8A phospho T72

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Publications (4)

Recent publications for all applications. Explore the full list and refine your search

The Journal of biological chemistry 299:105192 PubMed37625589

2023

Endogenous Rab38 regulates LRRK2's membrane recruitment and substrate Rab phosphorylation in melanocytes.

Applications

Unspecified application

Species

Unspecified reactive species

Alexandra Unapanta,Farbod Shavarebi,Jacob Porath,Yiyi Shen,Carson Balen,Albert Nguyen,Josh Tseng,Weng Si Leong,Michelle Liu,Pawel Lis,Santiago M Di Pietro,Annie Hiniker

Proceedings of the National Academy of Sciences of the United States of America 118: PubMed34088839

2021

Conformation and dynamics of the kinase domain drive subcellular location and activation of LRRK2.

Applications

Unspecified application

Species

Unspecified reactive species

Sven H Schmidt,Jui-Hung Weng,Phillip C Aoto,Daniela Boassa,Sebastian Mathea,Steve Silletti,Junru Hu,Maximilian Wallbott,Elizabeth A Komives,Stefan Knapp,Friedrich W Herberg,Susan S Taylor

Structure (London, England : 1993) 28:406-417.e6 PubMed32017888

2020

Structural Basis for Rab8a Recruitment of RILPL2 via LRRK2 Phosphorylation of Switch 2.

Applications

Unspecified application

Species

Unspecified reactive species

Dieter Waschbüsch,Elena Purlyte,Prosenjit Pal,Emma McGrath,Dario R Alessi,Amir R Khan

The Biochemical journal 475:1-22 PubMed29127256

2018

Development of phospho-specific Rab protein antibodies to monitor activity of the LRRK2 Parkinson's disease kinase.

Applications

Unspecified application

Species

Unspecified reactive species

Pawel Lis,Sophie Burel,Martin Steger,Matthias Mann,Fiona Brown,Federico Diez,Francesca Tonelli,Janice L Holton,Philip Winglok Ho,Shu-Leong Ho,Meng-Yun Chou,Nicole K Polinski,Terina N Martinez,Paul Davies,Dario R Alessi

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free

Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free (AB231706)

Dot Blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] - BSA and Azide free (AB231706)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Specificity

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Properties and storage information

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Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (4)

Product promise