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GAD67 Recombinant Antibody [PSH08-30] - Rabbit IgG (Chimeric)

Usd: 385

Specification

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Catalog# HA724362

GAD67 Recombinant Antibody [PSH08-30] - Rabbit IgG (Chimeric)

Application

  • WB

  • IHC-P

  • IF-Tissue

  • mIHC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

GAD67 Recombinant Antibody [PSH08-30] - Rabbit IgG (Chimeric)

Antibody Type

Recombinant Chimeric Antibody

Immunogen

Recombinant protein within human GAD67 aa 1-594.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Tissue, mIHC

Target Molecular Weight

Predicted band size: 67 kDa

Positive Control

Mouse brain tissue lysate, Rat brain tissue lysate.

Conjugation

unconjugated

Clone Number

PSH08-30

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P IHC-Fr IF-Tissue mIHC
human
 
Tested Tested
 
 
 
mouse
Tested Tested
Tested Tested
Not Recommended
Tested Tested
Tested Tested
rat
Tested Tested
Tested Tested
Not Recommended
Tested Tested
Tested Tested

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:1,000

  • IF-Tissue

  • 1:500

  • mIHC

  • 1:1,000

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.

UNIPROT

SwissProt: Q99259 Human

SwissProt: P48318 Mouse

SwissProt: P18088 Rat

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

Expand

Images

  • mIHC analysis of mouse cerebellum tissue (paraffin-embedded) with GAD67 (<a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000), QKI (<a href="/products/HA601377" style="font-weight: bold;text-decoration: underline;">HA601377</a>, 1/500) and CNPase (<a href="/products/ET1702-46" style="font-weight: bold;text-decoration: underline;">ET1702-46</a>, 1/2,000).<br /><br />The immunostaining was performed with the IRISKitCmTSA Kit (<a href="/products/900802" style="font-weight: bold;text-decoration: underline;">900802</a>). DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    mIHC analysis of mouse cerebellum tissue (paraffin-embedded) with GAD67 (HA724362, 1/1,000), QKI (HA601377, 1/500) and CNPase (ET1702-46, 1/2,000).

    The immunostaining was performed with the IRISKitCmTSA Kit (900802). DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

  • mIHC analysis of rat cerebellum tissue (paraffin-embedded) with GAD67 (<a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000), QKI (<a href="/products/HA601377" style="font-weight: bold;text-decoration: underline;">HA601377</a>, 1/500) and CNPase (<a href="/products/ET1702-46" style="font-weight: bold;text-decoration: underline;">ET1702-46</a>, 1/2,000).<br /><br />The immunostaining was performed with the IRISKitCmTSA Kit (<a href="/products/900802" style="font-weight: bold;text-decoration: underline;">900802</a>). DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    mIHC analysis of rat cerebellum tissue (paraffin-embedded) with GAD67 (HA724362, 1/1,000), QKI (HA601377, 1/500) and CNPase (ET1702-46, 1/2,000).

    The immunostaining was performed with the IRISKitCmTSA Kit (900802). DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

  • Application: Immunofluorescence (IF-tissue) <br /> <br />Species: Mouse <br />Tissue: Hippocampus <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× PBST <br />Endogenous peroxidase blocking: 3% H2O2, 10 minutes. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: GAD67, 1/500 (<a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, Rabbit, Green), VGLUT1, 1/500 (<a href="/products/HA601449" style="font-weight: bold;text-decoration: underline;">HA601449</a>, Guinea pig, Red), overnight at 4℃. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), Goat anti-Guinea pig IgG (iFluor&trade; 594, <a href="/products/HA1143" style="font-weight: bold;text-decoration: underline;">HA1143</a>), 1.5 hours at room temperature.

    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°C.

    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 (HA724362, Rabbit, Green), VGLUT1, 1/500 (HA601449, Guinea pig, Red), overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), Goat anti-Guinea pig IgG (iFluor™ 594, HA1143), 1.5 hours at room temperature.

  • Application: Immunofluorescence (IF-tissue) <br /> <br />Species: Rat <br />Tissue: Hippocampus <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× PBST <br />Endogenous peroxidase blocking: 3% H2O2, 10 minutes. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: GAD67, 1/500 (<a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, Rabbit, Green), VGLUT1, 1/500 (<a href="/products/HA601449" style="font-weight: bold;text-decoration: underline;">HA601449</a>, Guinea pig, Red), overnight at 4℃. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), Goat anti-Guinea pig IgG (iFluor&trade; 594, <a href="/products/HA1143" style="font-weight: bold;text-decoration: underline;">HA1143</a>), 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°C.

    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 (HA724362, Rabbit, Green), VGLUT1, 1/500 (HA601449, Guinea pig, Red), overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), Goat anti-Guinea pig IgG (iFluor™ 594, HA1143), 1.5 hours at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Cerebellum <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Cerebellum
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724362, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Cerebellum <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Cerebellum
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724362, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Cerebellum <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Cerebellum
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724362, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Hippocampus <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Hippocampus
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724362, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Hippocampus <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Hippocampus
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724362, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of GAD67 on different lysates with Rabbit anti-GAD67 antibody (<a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>) at 1/5,000 dilution. <br /> <br />Lane 1: Mouse brain tissue lysate <br />Lane 2: Mouse liver tissue lysate (negative) <br />Lane 3: Rat brain tissue lysate <br /> <br />Lysates/proteins at 20 µg/Lane. <br />Exposure time: 3 minutes; ECL: K1802 <br /> <br />Mouse liver is a negative control (PMID: 33968199). <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724362" style="font-weight: bold;text-decoration: underline;">HA724362</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 67 kDa <br />Observed band size: 67 kDa

    ☑ Relative expression (RE)

    Western blot analysis of GAD67 on different lysates with Rabbit anti-GAD67 antibody (HA724362) at 1/5,000 dilution.

    Lane 1: Mouse brain tissue lysate
    Lane 2: Mouse liver tissue lysate (negative)
    Lane 3: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 3 minutes; ECL: K1802

    Mouse liver is a negative control (PMID: 33968199).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724362, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 67 kDa
    Observed band size: 67 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"