Anti-vGAT-SAP [IT-71, KIT-71]

a tool for eliminating cells that express express the vesicular GABA transporter; targeted via affinity-purified anti-vGAT, eliminated via saporin

SKU: IT-71 Category: Quantity: Individual 25 ug, Individual 100 ug, Individual 250 ug, Individual 1 mg, Kit w/controls 25 ug, Kit w/controls 100 ug, Kit w/controls 250 ug | Antibody Type: affinity-purified, Polyclonal | Host: rabbit | Reactivity: guinea pig, human, mouse, pig, rat | Conjugate: streptavidin-saporin | Usage: eliminates cells, retrograde transport |

Vesicular GABA transporters (vGAT) mediate the accumulation of GABA into synaptic vesicles and its release from nerve terminals. This transporter is expressed in the nerve endings of GABAergic neurons throughout the CNS and has also been found in the pancreas and pituitary gland. During development, expression of the vGAT protein changes. Expression can also change in response to patterns of neuronal activity.

Anti-vGAT-SAP is a bonded toxin between a biotinylated rabbit polyclonal antibody and the secondary conjugate Streptavidin-ZAP containing the ribosome-inactivating protein, saporin. It specifically eliminates cells that express the vesicular GABA transporter.

Anti-vGAT-SAP is available individually (Cat. #IT-71) or as a kit (Cat. #KIT-71) which includes Anti-vGAT-SAP and BIgG-SAP Rabbit (Cat. #IT-75).

keywords: Vesicular GABA transporter, vGAT, Anti-vGAT, Anti-Vescular GABA Transporter, saporin, GABAergic neurons, brain, neuroscience

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Targeting vesicular gaba transporter (vGAT)-expressing cells with a polyclonal antibody to the lumenal domain of vGAT: results with a saporin conjugate.

Friedman CA, Russell BJ, Kohls MD, Ancheta LR, Shramm PA, Lappi DA (2016) Targeting vesicular gaba transporter (vGAT)-expressing cells with a polyclonal antibody to the lumenal domain of vGAT: results with a saporin conjugate. Neuroscience 2016 Abstracts 124.06 / E30. Society for Neuroscience, San Diego, CA. PMID: 0

Summary: The vesicular GABA transporter (vGAT) mediates the accumulation of GABA into synaptic vesicles and the release from these vesicles. vGAT is expressed in nerve endings of GABAergic neurons throughout the CNS. The GABAergic system is crucial for the development and functional maturation of the nervous system, as well as the maintenance of balance between excitation and inhibition required for normal neural circuit function. A panel of research tools has been created that target the lumenal domain of vGAT. Antiserum was raised against a peptide from the C-terminus of rat vGAT and resulted in an affinity-purified antibody and an immunotoxin specific for vGAT-expressing cells. The antigen sequence is identical among human, rat, mouse, pig and guinea pig. A stably-transfected clone of HEK293 cells (2E11HEK) that expresses vGAT on the cell surface shows excellent results for western blot, ICC and flow cytometry using both the antiserum and affinity-purified antibody. The affinity-purified antibody was used to create an immunotoxin by conjugating it to the ribosome-inactivating protein, saporin. Saporin irreversibly inactivates ribosomes, blocking protein synthesis, when it is escorted into a cell. Saporin cannot enter a cell on its own, but when escorted by something that binds to a cell surface marker it is internalized along with the binding moiety and causes cell death. The immunotoxin (Anti-vGAT-SAP) is 1000-fold more cytotoxic to 2E11HEK cells than non-conjugated saporin, based on the EC50 in a cytotoxicity assay. The affinity-purified vGAT antibody binds specifically to cells that express vGAT, and delivers a payload to the interior of these cells. Anti-vGAT-SAP could be an important tool in studying diseases involving dysfunction of GABAergic neurons. GABAergic neuron dysfunction is thought to be an underlying factor in Epilepsy, Down Syndrome, Fragile X Syndrome, Schizophrenia and Autism. In vivo, elimination of vGAT-expressing cells in a particular area (rather than knocking out vGAT systemically) makes it possible to study the functions of those regional cells. Animals can then be tested behaviorally before and after injections of Anti-vGAT-SAP to demonstrate the effects of loss of cells in a particular region of interest.

Related Products: vGAT Rabbit Polyclonal (Cat. #AB-N44), Anti-vGAT-SAP (Cat. #IT-71)

Oscillatory coupling within neonatal prefrontal-hippocampal networks is independent of selective removal of GABAergic neurons in the hippocampus.

Bitzenhofer SH, Hanganu-Opatz IL (2014) Oscillatory coupling within neonatal prefrontal-hippocampal networks is independent of selective removal of GABAergic neurons in the hippocampus. Neuropharmacology 77:57-67. doi: 10.1016/j.neuropharm.2013.09.007 PMID: 24056266

Summary: During cognitive tasks neuronal networks are entrained by oscillatory electrical rhythms with different frequencies. It has been proposed that GABAergic neurons in the prefrontal-hippocampal networks control this processing. The authors administered 252 ng of Anti-vGAT-SAP (Cat. #IT-71) into the ventral hippocampus of rats to examine how the GABAergic neurons could be involved. Unconjugated anti-vGAT (Cat #AB-N44) was used as a control. Hippocampal sharp waves were impaired during neonatal development, but the data indicate that oscillatory coupling between the neonatal prefrontal cortex and hippocampus is not controlled by GABAergic hippocampal interneurons.

Related Products: Anti-vGAT-SAP (Cat. #IT-71), vGAT Rabbit Polyclonal (Cat. #AB-N44)

Cracking down on inhibition: Selective removal of GABAergic interneurons from hippocampal networks.

Antonucci F, Alpár A, Kacza J, Caleo M, Verderio C, Giani A, Martens H, Chaudhry FA, Allegra M, Grosche J, Michalski D, Erck C, Hoffmann A, Harkany T, Matteoli M, Härtig W (2012) Cracking down on inhibition: Selective removal of GABAergic interneurons from hippocampal networks. J Neurosci 32(6):1989-2001. doi: 10.1523/JNEUROSCI.2720-11.2012

Related Products: Anti-vGAT-SAP (Cat. #IT-71)

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BIgG-SAP Rabbit (Cat. #IT-75)

vGAT Products:

vGAT Rabbit Polyclonal (Cat. #AB-N44) | affinity-purified (Cat. #AB-N44AP) | affinity-purified biotin-labeled (Cat. #AB-N44AP-BT) | affinity-purified Alexa488-labeled (Cat. #AB-N44AP-FLA)

Anti-vGAT-SAP (Cat. #IT-71)

Toolbox: vGAT Products

Cytotoxicity Assay for Targeted Toxins in vitro

Concentration Calculation Explained: Convert molarity to mg/ml and mg/ml to molarity

Preparing and Interpreting Cytotoxicity Data in vitro


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