VGAT (vesicular GABA transporter)

GABA transporter in the membrane of synaptic vesicles

Fact Sheet

Monoclonal mouse antibody, cytoplasmic domain
cytoplasmic, mouse -, rabbit -, guinea pig lumenal, rabbit
Cat. No. 131 011 IgG, unlabeled	100 µg purified IgG, lyophilized. For reconstitution add 100 µl H₂O to get a 1mg/ml solution of antibody in PBS. Then aliquot and store at -20°C until use.
Cat. No. 131 011C3 IgG, fluorescence-labeled	100 µg purified IgG, lyophilized, fluorescence-labeled with Oyster^® 550. Rabbit serum albumin was added for stabilization. For reconstitution add 100 µl H₂O to get a 1mg/ml solution of antibody in PBS. Add 1:1 (v/v) glycerol, then aliquot and store at -20°C until use. Reconstitute immediately upon receipt! Avoid bright light when working with the antibody to minimize photo bleeching of the fluorescent dye. The mounting agent Aquatex^® (Merck Chemicals) is not compatible with Oyster dyes!
Applications	WB: 1 : 500 up to 1 : 2000 (AP staining) blot IP: yes ICC: 1 : 200 up to 1 : 1000 131 011 131 011C3 IHC: 1 : 100 up to 1 : 1000 131 011
Clone	117G4
Subtype	IgG3
Immunogen	Synthetic peptide AEPPVEGDIHYQR (aa 75 - 87 in rat) coupled to key-hole limpet hemocyanin via an added N- terminal cysteine.
Reactivity	Human, rat, mouse, monkey; other species not tested yet.
Specificity	Specific for mammalian VGAT.
Remarks	VGAT aggregates after boiling, making it necessary to run SDS-PAGE only with non-boiled samples.

Polyclonal rabbit antibody, cytoplasmic domain
Cat. No. 131 002 crude antiserum	200 µl antiserum, lyophilized. For reconstitution add 200 µl H₂O, then aliquot and store at -20°C until use.
Cat. No. 131 003 purified antibody	50 µg specific antibody, lyophilized. Affinity purified with the immunogen. Rabbit serum albumin was added for stabilization. For reconstitution add 50 µl H₂O to get a 1mg/ml solution of antibody in PBS. Then aliquot and store at -20°C until use.
Applications	WB: 1 : 1000 up to 1 : 5000 (AP staining) 131 002 IP: yes ICC: 1 : 100 up to 1 : 1000 131 002 IHC: 1 : 500 up to 1 : 3000 131 002
Immunogen	Synthetic peptide AEPPVEGDIHYQR (aa 75 - 87 in rat) coupled to key-hole limpet hemocyanin via an added N- terminal cysteine.
Reactivity	Human, rat, mouse, monkey; other species not tested yet.
Specificity	Specific for VGAT.
Remarks	VGAT aggregates after boiling, making it necessary to run SDS-PAGE only with non-boiled samples.

Control peptide for cytoplasmic domain mouse and rabbit antibodies
Cat. No. 131-0P	100 µg lyophilized peptide. For reconstitution add 100 µl H₂O to get a 1mg/ml solution in PBS. Then aliquot and store at -20°C until use. Control peptides should also be stored at -20°C when still lyophilized!
Recommended dilution	Optimal concentrations should be determined by the end-user.
Remarks	This control peptide consists of the synthetic peptide (AEPPVEGDIHYQR) that has been used for immunization. It has been tested in preadsorption experiments and blocks efficiently and specifically the corresponding signal in Western blots. The amount of peptide needed for efficient blocking depends on the titer and on the affinity of the antibody to the antigen.

Polyclonal guinea pig antibody, cytoplasmic domain
Cat. No. 131 004 crude antiserum	100 µl antiserum, lyophilized. For reconstitution add 100 µl H₂O, then aliquot and store at -20°C until use.
Cat. No. 131 005 purified antibody	50 µg specific antibody, lyophilized. Affinity purified with the immunogen. Guinea pig serum albumin was added for stabilization. For reconstitution add 50 µl H2O to get a 1mg/ml solution of antibody in PBS. Then aliquot and store at -20°C until use.
Applications	WB: 1 : 1000 up to 1 : 2000 (AP staining) 131 004 131 005 IP: not tested yet ICC: 1 : 500 up to 1 : 1000 131 004 131 005 IHC: 1 : 200 up to 1 : 500 (cat. no. 131 004)
Immunogen	Strep-Tag^® fusion protein of rat VGAT (aa 2 - 115).
Reactivity	Rat, mouse; other species not tested yet.
Specificity	Specific for VGAT.
Remarks	VGAT aggregates after boiling, making it necessary to run SDS-PAGE only with non-boiled samples.

Polyclonal rabbit antibody, lumenal domain
Cat. No. 131 103 purified antibody	50 µg specific antibody, lyophilized. Affinity purified with the immunogen. Rabbit serum albumin was added for stabilization. For reconstitution add 50 µl H₂O to get a 1mg/ml solution of antibody in PBS. Then aliquot and store at -20°C until use.
Cat. No. 131 103C3 purified antibody, fluorescence-labeled	50 µg specific antibody, lyophilized. Affinity purified with the immunogen, fluorescence-labeled with Oyster^® 550. Rabbit serum albumin was added for stabilization. For reconstitution add 50 µl H₂O to get a 1mg/ml solution of antibody in PBS. Add 1:1 (v/v) glycerol, then aliquot and store at -20°C until use. Reconstitute immediately upon receipt! Avoid bright light when working with the antibody to minimize photo bleeching of the fluorescent dye. The mounting agent Aquatex^® (Merck Chemicals) is not compatible with Oyster dyes!
Applications	WB: 1 : 1000 (AP staining) IP: yes ICC: yes image IHC: yes
Immunogen	Synthetic peptide HSLEGLIEAYRTNAED (aa 510 - 525 in rat) coupled to key-hole limpet hemocyanin via an added N-terminal cysteine residue.
Reactivity	Human, rat, mouse; other species not tested yet.
Specificity	Specific for VGAT.
Remarks	VGAT aggregates after boiling, making it necessary to run SDS-PAGE only with non-boiled samples. These antibodies are intended for labeling of recycling synaptic vesicles at inhibitory nerve terminals by adding to living neurons. Further details see Martens et al. 2008.