RecombinantRabMAb

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

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Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)
  • Western blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)
  • Dot Blot - Anti-RAB8A (phospho T72) antibody [MJF-R20] (ab230260)
  • 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

Overview

  • 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.

Properties

  • 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

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Dot
Rat
WB
Mouse
Application Abreviews Notes
WB (2)
1/1000. Predicted molecular weight: 24 kDa.
Dot blot
1/1000.
Notes
WB
1/1000. Predicted molecular weight: 24 kDa.
Dot blot
1/1000.

Target

  • Function

    May 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 similarities

    Belongs to the small GTPase superfamily. Rab family.

  • Cellular localization

    Cell 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

    see all

  • Alternative names

    • AA409338 antibody
    • MEL antibody
    • Mel transforming oncogene (derived from cell line NK14) RAB8 homolog antibody
    • Mel transforming oncogene (derived from cell line NK14) antibody
    • Mel transforming oncogene (RAB8 homolog) antibody
    • Mel transforming oncogene antibody
    • MGC124948 antibody
    • Oncogene c mel antibody
    • Oncogene c-mel antibody
    • RAB8 antibody
    • RAB8A antibody
    • RAB8A member RAS oncogene family antibody
    • RAB8A_HUMAN antibody
    • Ras associated protein RAB8 antibody
    • Ras related protein Rab 8A antibody
    • Ras-related protein Rab-8A antibody
    see all

Images

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

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

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.

Click here to view the general protocols

References (4)

Publishing research using ab230260? Please let us know so that we can cite the reference in this datasheet.

ab230260 has been referenced in 4 publications.

  • Sanyal A  et al. LRRK2 Kinase Inhibition Rescues Deficits in Lysosome Function Due to Heterozygous GBA1 Expression in Human iPSC-Derived Neurons. Front Neurosci 14:442 (2020). PubMed: 32499675
  • Lara Ordónez AJ  et al. RAB8, RAB10 and RILPL1 contribute to both LRRK2 kinase-mediated centrosomal cohesion and ciliogenesis deficits. Hum Mol Genet 28:3552-3568 (2019). PubMed: 31428781
  • Berndsen K  et al. PPM1H phosphatase counteracts LRRK2 signaling by selectively dephosphorylating Rab proteins. Elife 8:N/A (2019). PubMed: 31663853
  • 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). PubMed: 29127256

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-RAB8A (phospho T72) antibody [MJF-R20]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Dopaminergic Neurons)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Dopaminergic Neurons
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Nov 10 2020

Western blot abreview for Anti-RAB8A (phospho T72) antibody [MJF-R20]

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (kidney)
Gel Running Conditions
Reduced Denaturing (4-12% bis-tris gel, MES buffer)
Loading amount
30 µg
Specification
kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Nov 03 2020

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