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
  • Host species
    Rabbit
  • 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

    Synthetic peptide within Human GABARAP+GABARAPL1+GABARAPL2 aa 1-100. The exact sequence is proprietary.

  • Positive control
    • Fetal kidney lysate, fetal brain lysate, SH-SY5Y cell lysate, HepG2 whole cell lysate (ab7900), 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.

Ssee IHC antigen retrieval protocols.

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.
  • Target

    • Relevance
      GABARAP: Ubiquitin-like modifier that plays a role in intracellular transport of GABA(A) receptors and its interaction with the cytoskeleton. Involved in apoptosis. Involved in autophagy. Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation.
      GABARAPL1: Ubiquitin-like modifier that increases cell-surface expression of kappa-type opioid receptor through facilitating anterograde intracellular trafficking of the receptor. Involved in formation of autophagosomal vacuoles. Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation.
      GABARAPL2: Ubiquitin-like modifier involved in intra-Golgi traffic. Modulates intra-Golgi transport through coupling between NSF activity and SNAREs activation. It first stimulates the ATPase activity of NSF which in turn stimulates the association with GOSR1 (By similarity). Involved in autophagy. Plays a role in mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation.
    • Database links

    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
      All lanes : 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.

    References

    This product has been referenced in:
    • Cackovic J  et al. Vulnerable Parkin Loss-of-FunctionDrosophilaDopaminergic Neurons Have Advanced Mitochondrial Aging, Mitochondrial Network Loss and Transiently Reduced Autophagosome Recruitment. Front Cell Neurosci 12:39 (2018). Read more (PubMed: 29497364) »
    • De Pace R  et al. Altered distribution of ATG9A and accumulation of axonal aggregates in neurons from a mouse model of AP-4 deficiency syndrome. PLoS Genet 14:e1007363 (2018). Read more (PubMed: 29698489) »
    See all 15 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Q&A

    Answer

    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 92%. This means that a cross-reactivity is very likely. In addition, this antibody has been established in both WB and IHC.


    However, to our knowledge, ab109364 has not been tested in silkworm . However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

    Simply follow these easy steps below to apply for our AbTrial Program:

    1. Reply to this email, letting us know you are interested in testing this product.

    2. Our scientists will email you an inactive personal discount code for the value of the product.

    3. Purchase and test the product at the regular price.

    4. Submit your results, including your discount code in the additional notes section of your Abreview.

    5. Once the Abreview is submitted, the discount code will become active.

    6. Apply your discount code on your next order to receive that value off.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

    Read More

    Answer

    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 code has an expiry date of 4 month after issuing. So issuing it now would eat up your time for experiments unnecessarily.

    If you write us an email when you know you would like to use it, we generally answer within 24 hours. This means there will be no additional delay for you then.

    Read More

    Answer

    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 more.

    I checked our other Anti-GABARAP antibodies and would like to recommend ab109364 (https://www.abcam.com/index.html?datasheet=109364 (or use the following: https://www.abcam.com/index.html?datasheet=109364).). The immunogen shows 92% homology with Drosophila melanogaster and therefore it is likely to cross react.

    To our knowledge, ab109364 has not been experimentally tested in Drosophila melanogaster. Therefore, I can offer a discount off a future purchase if you buy ab109364 now, test it in Drosophila melanogaster and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of ab109364 off a future order.

    If you are interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that you would like to proceed and test ab109364 in Drosophila melanogaster. I will then send a discount code. This code must be issued before purchasing ab109364 so please wait for my reply before ordering.

    2. Purchase ab109364 either by phone, fax, or online (www.abcam.com).

    3. Test it in Drosophila melanogaster.

    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyproduct ordered and the discount code is valid for 4 months after issue.
    Please remember that submission of the Abreview is sufficient for the discount code to become active.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab109364 turns out to be unsuitable for Drosophila melanogaster, you will still receive the discount on your next purchase after your Abreview has been submitted.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

    Read More

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

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