Recombinant
RabMAb

Recombinant Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20842] to ABAT/GABA-T
  • Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

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. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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

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

    see all

  • 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

  • Western blot - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Western blot - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunocytochemistry/ Immunofluorescence - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunocytochemistry/ Immunofluorescence - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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.

  • Western blot - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Western blot - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    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).

  • Immunoprecipitation - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunoprecipitation - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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 Cytometry - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Flow Cytometry - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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 Cytometry - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Flow Cytometry - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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.

  • Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Frozen sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABAT/GABA-T antibody [EPR20842] (ab216465)

    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).

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (1)

Publishing research using ab216465? Please let us know so that we can cite the reference in this datasheet.

ab216465 has been referenced in 1 publication.

  • 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). PubMed: 30535457

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