Recombinant
RabMAb

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

Rabbit recombinant monoclonal RAB8A (phospho T72) antibody [MJF-R20]. Validated in WB, Dot and tested in Mouse, Human. Cited in 1 publication(s).

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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

    Suitable for: WB, Dot blotmore details

  • Species reactivity

    Reacts 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.

     

    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

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    MJF-R20

  • Isotype

    IgG

  • Research areas

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

  • 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

    • 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.

    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) »
    See 1 Publication for this product

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

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