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

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, Flow Cyt (Intra), WB, IHC-P, IHC-Fr, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20842
Isotype

IgG
Research areas

Alternative Versions
- Anti-ABAT/GABA-T antibody [EPR20842] - BSA and Azide free (ab233702)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human ABAT/GABA-T protein (ab152164)
Related Products
- Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

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.
Flow Cyt (Intra)		1/60.
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.
IP		1/30.

Notes
ICC/IF 1/100.
Flow Cyt (Intra) 1/60.
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.
IP 1/30.

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.
Target information above from: UniProt accession P80404 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 18 Human
- Entrez Gene: 268860 Mouse
- Entrez Gene: 81632 Rat
- Omim: 137150 Human
- SwissProt: P80404 Human
- SwissProt: P61922 Mouse
- SwissProt: P50554 Rat
- Unigene: 336768 Human
- Unigene: 259315 Mouse
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

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)

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)

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)

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)

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

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

Intracellular 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 (cellsincubated 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)

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)

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)

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)

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)

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)

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)

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)

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

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

SDS download

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Datasheet download

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

ab216465 has been referenced in 2 publications.

Zheng J et al. Interneuron Accumulation of Phosphorylated tau Impairs Adult Hippocampal Neurogenesis by Suppressing GABAergic Transmission. Cell Stem Cell 26:331-345.e6 (2020). PubMed: 31978364
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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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As