Overview

  • Product name

    Anti-ABAT/GABA-T antibody [EPR20842]
    See all ABAT/GABA-T primary antibodies
  • Description

    Rabbit monoclonal [EPR20842] to ABAT/GABA-T
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ABAT/GABA-T aa 250-350. The exact sequence is proprietary.
    Database link: P80404

  • Positive control

    • WB: MCF7, HepG2 and SH-SY5Y whole cell lysate; Human brain and fetal liver lysate; Mouse brain and liver lysate; Rat liver and brain lysate. IHC-P: Human kidney and breast cancer tissue; Mouse testis tissue; Rat liver tissue. IHC-Fr: Mouse cerebellum and liver tissue; Mouse and rat choroid plexus tissue. ICC/IF: MCF7 and HepG2 cells. Flow: HepG2 and MCF7 cells. IP: MCF7 whole cell lysate.
  • General notes

    Previously labelled as ABAT. 

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab216465 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
ICC/IF 1/100.
WB 1/1000. Predicted molecular weight: 56 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/500.
Flow Cyt 1/60.
IP 1/30.

Target

  • Function

    Catalyzes the conversion of gamma-aminobutyrate and L-beta-aminoisobutyrate to succinate semialdehyde and methylmalonate semialdehyde, respectively. Can also convert delta-aminovalerate and beta-alanine.
  • Tissue specificity

    Liver > pancreas > brain > kidney > heart > placenta.
  • Involvement in disease

    Defects in ABAT are a cause of gamma-aminobutyrate transaminase deficiency (GABA-AT deficiency) [MIM:613163]. The phenotype of this deficiency includes psychomotor retardation, hypotonia, hyperreflexia, lethargy, refractory seizures, and EEG abnormalities.
  • Sequence similarities

    Belongs to the class-III pyridoxal-phosphate-dependent aminotransferase family.
  • Cellular localization

    Mitochondrion matrix.
  • Information by UniProt
  • Database links

  • Alternative names

    • (S) 3 amino 2 methylpropionate transaminase antibody
    • (S)-3-amino-2-methylpropionate transaminase antibody
    • 4 aminobutyrate aminotransferase antibody
    • 4 aminobutyrate aminotransferase, mitochondrial antibody
    • 4-aminobutyrate aminotransferase antibody
    • ABAT antibody
    • FLJ17813 antibody
    • FLJ30272 antibody
    • GABA aminotransferase antibody
    • GABA AT antibody
    • GABA T antibody
    • GABA transaminase antibody
    • GABA transferase antibody
    • GABA-AT antibody
    • GABA-T antibody
    • GABAT antibody
    • GABT_HUMAN antibody
    • Gamma amino N butyrate transaminase antibody
    • Gamma-amino-N-butyrate transaminase antibody
    • hCG1984265 antibody
    • L AIBAT antibody
    • L-AIBAT antibody
    • LAIBAT antibody
    • mitochondrial antibody
    • NPD009 antibody
    see all

Images

  • All lanes : Anti-ABAT/GABA-T antibody [EPR20842] (ab216465) at 1/1000 dilution

    Lane 1 : Mouse liver lysate
    Lane 2 : Rat liver lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Human fetal liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
    Lane 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 4 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 56 kDa



    Blocking/diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: Lanes 1 and 2: 3 seconds; Lane 3: 5 seconds; Lane 4: 26 seconds.

    The observed molecular mass is consistent with what has been described in the literature (PMID: 11459221).

  • Immunofluorescent analysis of 100% methanol-fixed HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling ABAT/GABA-T with ab216465 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 (green). Confocal image showing mitochondrial staining in HepG2 cell line is observed. The nuclear counter stain is DAPI (blue).

    Mitochondria are stained with ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary both at 1/1000 dilution (red).

    -ve control 1: ab216465 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

    -ve control 2: ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker at 1/1000 dilution followed by ab150077 (AlexaFluor®488 Goat anti-Rabbit secondary) at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed MCF7 (human breast adenocarcinoma epithelial cell) cells labeling ABAT/GABA-T with ab216465 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 (green). Confocal image showing mitochondrial staining in MCF7 cell line. The nuclear counter stain is DAPI (blue).

    Mitochondria are stained with ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary both at 1/1000 dilution (red).

    -ve control 1: ab216465 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

    -ve control 2: ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker at 1/1000 dilution followed by ab150077 (AlexaFluor®488 Goat anti-Rabbit secondary) at 1/1000 dilution.

  • All lanes : Anti-ABAT/GABA-T antibody [EPR20842] (ab216465) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Human brain lysate
    Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 5 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 56 kDa



    Blocking/diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: Lane 1,2,3 and 4: 6 seconds; Lane 5: 92 seconds.

    The observed molecular mass is consistent with what has been described in the literature (PMID: 11459221).

  • ABAT/GABA-T was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate with ab216465 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216465 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution
    Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg (Input).
    Lane 2: ab216465 IP in MCF7 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216465 in MCF7 whole cell lysate (-).


    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

     

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling ABAT/GABA-T with ab216465 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabelled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cell line labeling ABAT/GABA-T with ab216465 at 1/60 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabelled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Cytoplasmic staining in rat liver is observed. Counter stained with DAPI (blue).
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat choroid plexus tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Cytoplasmic staining in rat choroid plexus (PMID:25239459, PMID: 11459221) is observed. Counter stained with DAPI (blue).
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse choroid plexus tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Cytoplasmic staining in mouse choroid plexus (PMID:25239459, PMID: 11459221) is observed. Counter stained with DAPI (blue).
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Cytoplasmic staining in mouse cerebellum (PMID:25239459, PMID: 11459221) is observed. Counter stained with DAPI (blue).
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in rat liver (PMID: 25771305; PMID: 25738457) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in mouse testis (PMID: 25771305; PMID: 25738457) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human breast cancer (PMID: 25771305; PMID: 25738457) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling ABAT/GABA-T with ab216465 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human kidney (PMID: 25771305; PMID: 25738457) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

References

This product has been referenced in:

  • Zhao G  et al. High methylation of the 4-aminobutyrate aminotransferase gene predicts a poor prognosis in patients with myelodysplastic syndrome. Int J Oncol 54:491-504 (2019). Read more (PubMed: 30535457) »
See 1 Publication for this product

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