Recombinant
RabMAb

Recombinant Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] - BSA and Azide free (ab225535)

Overview

  • Product name

    Anti-GABARAP+GABARAPL1+GABARAPL2 antibody [EPR4805] - BSA and Azide free
    See all GABARAP+GABARAPL1+GABARAPL2 primary antibodies
  • Description

    Rabbit monoclonal [EPR4805] to GABARAP+GABARAPL1+GABARAPL2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Silk worm
  • Immunogen

    Synthetic peptide aa 1-100. The exact sequence is proprietary.

  • Positive control

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

    Ab225535 is the carrier-free version of ab109364. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab225535 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.??

    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. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4805
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab225535 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: 14 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See 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.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
  • 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

    • 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109364).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109364).

    • 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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109364).

    • Unpurified ab109364, at 1/500, staining GABARAP+GABARAPL1+GABARAPL2 in Human brain tissue by immunohistochemistry.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109364).

    • This ICC data was generated using the same anti-GABARAP+GABARAPL1/2 antibody clone [EPR4805] in a different buffer formulation (cat# ab109364).

      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).

    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: (2017). ICC/IF ; Human . Read more (PubMed: 28195531) »
    • Cappelletti C  et al. Autophagy, inflammation and innate immunity in inflammatory myopathies. PLoS One 9:e111490 (2014). Radioimmunoprecipitation ; Human . Read more (PubMed: 25365350) »
    See all 2 Publications for this product

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    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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