GAD67 Recombinant Mouse Monoclonal Antibody [PSH08-30]
Overview
Product Name
GAD67 Recombinant Mouse Monoclonal Antibody [PSH08-30]
Antibody Type
Recombinant Mouse Monoclonal Antibody
Immunogen
Recombinant protein within human GAD67 aa 1-594.
Species Reactivity
Mouse, Rat
Validated Applications
WB, IHC-P, IHC-Fr, IF-Tissue
Molecular Weight
Predicted band size: 67 kDa
Positive Control
Mouse brain tissue lysate, Rat brain tissue lysate, mouse cerebellum tissue, rat cerebellum tissue.
Conjugation
unconjugated
Clone Number
PSH08-30
Product Features
Form
Liquid
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:10,000
-
IHC-P
-
1:500-1:2,000
-
IHC-Fr
-
1:500
-
IF-Tissue
-
1:500
Target
Function
There are two forms of glutamic acid decarboxylases (GADs) that are found in the brain: GAD-65 (also known as GAD2) and GAD-67 (also known as GAD1, GAD or SCP). GAD-65 and GAD-67 are members of the group II decarboxylase family of proteins and are responsible for catalyzing the rate limiting step in the production of GABA (g-aminobutyric acid) from L-glutamic acid. Although both GADs are found in the brain, GAD-65 localizes to synaptic vesicle membranes in nerve terminals, while GAD-67 is distributed throughout the cell. GAD-67 is responsible for the basal levels of GABA synthesis. In the case of a heightened demand for GABA in neurotransmission, GAD-65 will transiently activate to assist in GABA production. The loss of GAD-65 is detrimental and can impair GABA neurotransmission, however the loss of GAD-67 is lethal. Due to alternative splicing, two isoforms exist for GAD-67, the predominant GAD-67 form and the minor GAD-25 form. GAD-25 is not expressed in brain but can be found in a variety of endocrine tissues.
Background References
1. Li JT et al. Repeated Blockade of NMDA Receptors During Adolescence Impairs Reversal Learning and Disrupts GABAergic Interneurons in Rat Medial Prefrontal Cortex. Front Mol Neurosci 9:17 (2016).
2. Fu Q et al. MHC-I promotes apoptosis of GABAergic interneurons in the spinal dorsal horn and contributes to cancer induced bone pain. Exp Neurol 286:12-20 (2016).
Subcellular Location
Cytoplasm, Plasma Membrane.
Synonyms
67 kDa glutamic acid decarboxylase antibody
CPSQ1 antibody
DCE1 antibody
DCE1_HUMAN antibody
EC 4.1.1.15 antibody
FLJ45882 antibody
GAD 67 antibody
GAD antibody
GAD-67 antibody
GAD1 antibody
ExpandImages
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Immunofluorescence analysis of frozen mouse cerebellum tissue with Mouse anti-GAD67 antibody (HA601370) at 1/500 dilution.
Important Notice: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.
The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA601370, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/500 dilution. Nuclei were counterstained with DAPI (blue). -
Immunofluorescence analysis of frozen rat cerebellum tissue with Mouse anti-GAD67 antibody (HA601370) at 1/500 dilution.
Important Notice: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.
The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA601370, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/500 dilution. Nuclei were counterstained with DAPI (blue). -
Western blot analysis of GAD67 on different lysates with Mouse anti-GAD67 antibody (HA601370) at 1/10,000 dilution.
Lane 1: Mouse brain tissue lysate
Lane 2: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 67 kDa
Observed band size: 67 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601370) at 1/10,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Mouse anti-GAD67 antibody (HA601370) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601370) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Mouse anti-GAD67 antibody (HA601370) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601370) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Mouse anti-GAD67 antibody (HA601370) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601370) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue with Mouse anti-GAD67 antibody (HA601370) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601370) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue with Mouse anti-GAD67 antibody (HA601370) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601370) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue with Mouse anti-GAD67 antibody (HA601370) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601370) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Hippocampus
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× PBST
Endogenous peroxidase blocking: 3% H2O2, 10 minutes.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: GAD67, 1/500 (HA601370, Mouse, Green), VGLUT1, 1:500 (HA601449, Guinea pig, Red), overnight at 4℃.
Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), Goat anti-Guinea pig IgG (iFluor™ 594, HA1143), 1.5 hours at room temperature. -
Application: Immunofluorescence (IF-tissue)
Species: Rat
Tissue: Hippocampus
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× PBST
Endogenous peroxidase blocking: 3% H2O2, 10 minutes.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: GAD67, 1/500 (HA601370, Mouse, Green), VGLUT1, 1:500 (HA601449, Guinea pig, Red), overnight at 4℃.
Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), Goat anti-Guinea pig IgG (iFluor™ 594, HA1143), 1.5 hours at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"