Recombinant
RabMAb

Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364)

Overview

  • Product name
    Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805]
    See all GABARAP+GABARAPL1+GABARAPL2 primary antibodies
  • Description
    Rabbit monoclonal [EPR4805] to GABARAP+GABARAPL1+GABARAPL2
  • Specificity
    ab109364 will also recognize related targets GABARAPL1 and GABARAPL2: GABARAPL1: pBLAST 100% immunogen homology AND Abreview 38385 WB data GABARAPL2: pBLAST 93% homology (only 1aa gap of 14aa) AND Abreview 38385 WB data
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Silk worm
  • Immunogen

    corresponding to Human GABARAP+GABARAPL1+GABARAPL2 aa 1-100.

  • Positive control
    • Fetal kidney lysate, fetal brain lysate, SH-SY5Y cell lysate, HepG2 cell lysate, human brain tissue, HeLa cells.
  • General notes

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109364 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
Flow Cyt 1/20.
WB 1/1000 - 1/10000. Predicted molecular weight: 14 kDa.
IHC-P 1/500 - 1/1000.

Antigen retrieval is recommended: see protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

ICC/IF 1/100 - 1/500.
  • Application notes
    Is unsuitable for IP.
  • Images

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling GABARAP+GABARAPL1+GABARAPL2 with purified ab109364 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling GABARAP+GABARAPL1+GABARAPL2 with Purified ab109364 at 1:500 dilution (0.29 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GABARAP+GABARAPL1+GABARAPL2 with purified ab109364 at 1:20 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364) at 1/5000 dilution (purified) + Rat kidney lysates at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 14 kDa
      Observed band size : 17 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST

    • All lanes : Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364) at 1/5000 dilution (purified)

      Lane 1 : Mouse kidney lysates
      Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 14 kDa
      Observed band size : 17 kDa (why is the actual band size different from the predicted?)

      Blocking and diluting buffer: 5% NFDM/TBST

    • Unpurified ab109364 staining GABARAP+GABARAPL1+GABARAPL2 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black)

      Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    • All lanes : Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] (ab109364) at 1/1000 dilution (unpurified)

      Lane 1 : Fetal kidney lysate
      Lane 2 : SH-SY5Y cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : Fetal brain lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 14 kDa
    • Unpurified ab109364, at 1/500, staining GABARAP+GABARAPL1+GABARAPL2 in Human brain tissue by immunohistochemistry.

    • Unpurified ab109364, at 1/100, staining HeLa cells by immunofluorescence.

    References

    This product has been referenced in:
    • Jung J  et al. Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator. Elife 6:N/A (2017). ICC/IF ; Human . Read more (PubMed: 28195531) »
    • Romanelli D  et al. Roles and regulation of autophagy and apoptosis in the remodelling of the lepidopteran midgut epithelium during metamorphosis. Sci Rep 6:32939 (2016). Read more (PubMed: 27609527) »

    See all 4 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HepG2 cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    HepG2 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 13 2017

    Application
    Western blot
    Sample
    Human Tissue lysate - whole (skeletal muscle tissue)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    20 µg
    Specification
    skeletal muscle tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Aug 12 2015

    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Reduced Denaturing (12%)
    Sample
    Silk worm (Bombyx mori) Tissue lysate - whole (Midgut)
    Specification
    Midgut
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 02 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (13%)
    Sample
    Human Cell lysate - whole cell (SH-SY5Y cells stably expressing GFP, GFP-GABARAP,)
    Specification
    SH-SY5Y cells stably expressing GFP, GFP-GABARAP,
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Dr. George Allen

    Verified customer

    Submitted Sep 26 2013

    Application
    IHC - Wholemount
    Sample
    Fruit fly (Drosophila melanogaster) Tissue (fat body)
    Specification
    fat body
    Username

    Abcam user community

    Verified customer

    Submitted Sep 19 2013

    Thank you very much for your interest our AbTrial program.



    I performed a comparison of the immunogens for our GABARAP antibodies, and the identity between the immunogen of ab109364 and the silkworm protein looks very promising with ...

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    Thank you for your reply.

    I wish you good luck with the live reporter experiments.

    I will be happy to issue the discount code for you now, but I think it would be better to do this when you know that you will use it. The discount co...

    Read More

    Thank you very much for your interest in our antibodies.

    According to a quick blast check the immunogen of ab1398 has a homology with Drosophila melanogaster of 82%. We generally expect cross reaction of an antibody from 85% homology and mor...

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