Anti-RAB27A antibody [EPR3021] (ab108983)

Overview

  • Product nameAnti-RAB27A antibody [EPR3021]
    See all RAB27A primary antibodies
  • Description
    Rabbit monoclonal [EPR3021] to RAB27A
  • Tested applicationsSuitable for: WB, IP, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Human, Rhesus monkey
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human RAB27A aa 50-150.
    Database link: P51159

  • Positive control
    • WB: HeLa, MCF-7, Jurkat and HL-60 cell lysates. ICC/IF: Jurkat cells. Flow Cyt: THP-1 cells. IP: MCF-7 cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels.

    If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:

    Anti-RAB27A antibody (Alexa Fluor® 488) [EPR3021] (ab197900)

    Anti-RAB27A antibody (Alexa Fluor® 647) [EPR3021] (ab198601)

Properties

Applications

Our Abpromise guarantee covers the use of ab108983 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 1/5000. Predicted molecular weight: 25 kDa.

For unpurified use at 1/1000 - 1/10000.

IP 1/30.

For unpurified use at 1/10 - 1/100.

Flow Cyt 1/400.

For unpurified use at 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.
  • Application notesIs unsuitable for IHC-P.
  • Target

    • FunctionPlays a role in cytotoxic granule exocytosis in lymphocytes. Required for both granule maturation and granule docking and priming at the immunologic synapse.
    • Tissue specificityFound in all the examined tissues except in brain. Low expression was found in thymus, kidney, muscle and placenta. Detected in melanocytes, and in most tumor cell lines examined. Expressed in cytotoxic T-lymphocytes (CTL) and mast cells.
    • Involvement in diseaseDefects in RAB27A are a cause of Griscelli syndrome type 2 (GS2) [MIM:607624]. Griscelli syndrome is a rare autosomal recessive disorder that results in pigmentary dilution of the skin and hair, the presence of large clumps of pigment in hair shafts, and an accumulation of melanosomes in melanocytes. GS2 patients also develop an uncontrolled T-lymphocyte and macrophage activation syndrome, known as hemophagocytic syndrome, leading to death in the absence of bone marrow transplantation. Neurological impairment is present in some patients, likely as a result of hemophagocytic syndrome.
    • Sequence similaritiesBelongs to the small GTPase superfamily. Rab family.
    • Cellular localizationMembrane. Melanosome. Late endosome. Lysosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Localizes to endosomal exocytic vesicles.
    • Information by UniProt
    • Database links
    • Alternative names
      • GS2 antibody
      • GTP-binding protein Ram antibody
      • HsT18676 antibody
      • MGC117246 antibody
      • Mutant Ras related protein Rab-27A antibody
      • Rab-27 antibody
      • RAB-27A antibody
      • RAB27 antibody
      • RAB27A antibody
      • RAB27A member RAS oncogene family antibody
      • RAM antibody
      • Ras-related protein Rab-27A antibody
      • Ras-related protein Rab27A antibody
      • RB27A_HUMAN antibody
      see all

    Anti-RAB27A antibody [EPR3021] images

    • All lanes : Anti-RAB27A antibody [EPR3021] (ab108983) at 1/5000 dilution (purified)

      Lane 1 : MCF-7 cell lysate
      Lane 2 : Jurkat cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 25 kDa
      Observed band size : 28 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-RAB27A antibody [EPR3021] (ab108983) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa cell lysate
      Lane 2 : MCF-7 cell lysate
      Lane 3 : HL-60 cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 25 kDa
    • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling RAB27A with purified ab108983 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    • Unpurified ab108983 staining RAB27A in Rhesus monkey fetal kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 10% serum for 1 hour at 37°C. Samples were incubated with primary antibody (1/100 in 1% FCS/PBS) for 1 hour at 37°C. An Alexa Fluor® 568-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

      See Abreview

    • ab108983 (purified) at 1/30 immunoprecipitating RAB27A in MCF7 whole cell lysate (Lane 1). Lane 2 - Rabbit monoclonal IgG (ab172730) instead of ab81292 in MCF7 whole cell lysate. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • ab108983 (purified) at 1/30 immunoprecipitating RAB27A in HeLa whole cell lysate, 10ug (Lane 1). Lane 2 - Rabbit monoclonal IgG (ab172730) instead of ab81292 in HeLa whole cell lysate. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Flow Cytometry analysis of THP-1 cells labelling RAB27A with purified ab108983 at 1/400 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    References for Anti-RAB27A antibody [EPR3021] (ab108983)

    ab108983 has not yet been referenced specifically in any publications.

    Product Wall

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate, pH6, 100C, 20min
    Sample Human Tissue sections (tonsil, appendix)
    Specification tonsil, appendix
    Permeabilization No
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jan 27 2015

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
    Sample Rhesus monkey Cell (Fetal Rhesus Monkey Kidney Cell (FRhK-4 cell))
    Specification Fetal Rhesus Monkey Kidney Cell (FRhK-4 cell)
    Permeabilization No
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 02 2013

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"