RecombinantRabMAb

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

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

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) - BSA and Azide free
  • Suitable for: WB, Dot blot
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [MJF-R20] to RAB8A (phospho T72) - BSA and Azide free

  • Host species

    Rabbit

  • 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.

  • Tested applications

    Suitable for: WB, Dot blotmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HEK-293 cells transfected with LRRK2 [Y1699C] and HA-tagged Rab8A, Rab10 and Rab35 expression vectors, treated with 150 nM MLi-2.

  • General notes

    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).

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    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.

    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.

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab231706 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
Use at an assay dependent concentration. Predicted molecular weight: 24 kDa.
Dot blot
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 24 kDa.
Dot blot
Use at an assay dependent concentration.

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

  • Dot 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)

    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).

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

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

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

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 (3)

Publishing research using ab231706? Please let us know so that we can cite the reference in this datasheet.

ab231706 has been referenced in 3 publications.

  • Schmidt SH  et al. Conformation and dynamics of the kinase domain drive subcellular location and activation of LRRK2. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 34088839
  • Waschbüsch D  et al. Structural Basis for Rab8a Recruitment of RILPL2 via LRRK2 Phosphorylation of Switch 2. Structure 28:406-417.e6 (2020). PubMed: 32017888
  • 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

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