GAD1/GAD67 Antibody

R&D Systems | Catalog # AF2086

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Monkey, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all GAD1/GAD67 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human GAD1/GAD67
Ala2-Asn97
Accession # Q99259

Specificity

Detects human GAD1/GAD67 in direct ELISAs and detects human, mouse, and rat GAD1/GAD67 in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for GAD1/GAD67 Antibody

Detection of Human, Mouse, and Rat GAD1/GAD67 antibody by Western Blot.

Detection of Human, Mouse, and Rat GAD1/GAD67 by Western Blot.

Western blot shows lysates of human brain (hypothalamus) tissue, mouse brain (total) tissue, and rat brain (total) tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat GAD1/GAD67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2086) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody HAF017). A specific band was detected for GAD1/GAD67 at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
GAD1/GAD67 antibody in Human Spinal Cord by Immunohistochemistry (IHC-P).

GAD1/GAD67 in Human Spinal Cord.

GAD1/GAD67 was detected in immersion fixed paraffin-embedded sections of human spinal cord using 15 µg/mL Goat Anti-Human/Mouse/Rat GAD1/GAD67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2086) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Mouse GAD1/GAD67 antibody by Simple WesternTM.

Detection of Mouse GAD1/GAD67 by Simple WesternTM.

Simple Western lane view shows lysates of mouse brain tissue, loaded at 0.2 mg/mL. A specific band was detected for GAD1/GAD67 at approximately 65 kDa (as indicated) using 1 µg/mL of Goat Anti-Human/Mouse/Rat GAD1/GAD67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2086) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of GAD1/GAD67 by Immunohistochemistry

Detection of GAD1/GAD67 by Immunohistochemistry

Optogenetic inhibition of IPN GABAergic terminals in LDTg. A, Surgery and electrophysiology schematic illustrating cre-mediated expression of NpHR in GAD2-cre mice. B, Viral vector cre-mediated expression of NpHR (green) overlapping (yellow) with immunohistochemistry staining of GAD1/2 (red) in the IPN of GAD2-cre mice. C, Light evoked hyperpolarization of membrane potential in ex vivo recording from an IPN neuron expressing eYFP-tagged NpHR. D, Light evoked inhibitory outward current in ex vivo recording from eYFP-labeled IPN neuron expressing NpHR (Vm = −70 mV). E, Histologic confirmation of viral injection and eYFP-NpHR localization within the IPN. F, Surgery schematic illustrating cre-mediated expression of NpHR or eYFP in IPN of GAD2-cre mice and fiber-optic placement above the LDTg for light-evoked inhibition of terminals. G, Histologic confirmation of fiber-optic above the LDTg with eYFP-NpHR fluorescence in IPN neuron axon terminals. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40659528), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GAD1/GAD67 by Immunohistochemistry

Detection of GAD1/GAD67 by Immunohistochemistry

GABAergic IPN neurons synapse onto both GABAergic and glutamatergic LDTg neurons. A, Surgery schematic of rabies backlabeling of IPN neurons synapsing to LDTg subtypes using GAD2-Cre (N = 3), ChAT-Cre (N = 3), and Vglut2-Cre (N = 4) mice. B, GABAergic LDTg starter cells identified by overlap (orange) of helper virus (green) with rabies virus (red). White arrowhead points to representative starter cells identified within LDTg within a GAD2-cre mouse. Rabies only neurons (red alone) indicate local connections within LDTg to starter cells. C, Cre-dependent helper virus expression is restricted by using cre mouse lines. Immunohistochemistry labeling GAD67 (purple) overlaps (light blue) with cre-dependent helper virus (green) in GAD2-cre mouse line (top panels). GAD67 (purple) does not overlap with helper virus (green) in Vglut2-cre mouse line. D, Total rabies positive cells in LDTg divided into proportion of starter cells (rabies+, helper virus+) and local LDTg connections (rabies+, helper virus−). Rabies virus cells were identified in Chat-Cre, GAD2-cre, and Vglut2-cre mouse lines. E, IPN rabies+ neurons are present in multiple IPN subregions. F, IPN rabies+ neurons are identified as GAD+ or GAD− using GAD67 immunohistochemistry. G, IPN inputs proportion to starter cells show IPN connections to all three LDTg subtypes (cholinergic, GABAergic, and glutamatergic) but shows more connections to GABAergic and glutamatergic LDTg neurons. H, IPN subregions may show different patterns of distribution to LDTg subtypes. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40659528), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for GAD1/GAD67 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human spinal cord

Simple Western

1 µg/mL
Sample:   Mouse brain tissue

Western Blot

1 µg/mL
Sample:   Human brain (hypothalamus) tissue, mouse brain (total) tissue, and rat brain (total) tissue

Reviewed Applications

Read 1 review rated 1 using AF2086 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: GAD1/GAD67

GAD1, also named 67 kDa or brain GAD, is an enzyme that catalyzes the formation of the inhibitory neurotransmitter gamma -amino butyric acid (GABA) from glutamate. GAD1 is also expressed in multiple non-neuronal tissues during development.

Long Name

Glutamate Decarboxylase 1 (Brain, 67kDa)

Alternate Names

GAD67

Entrez Gene IDs

2571 (Human)

Gene Symbol

GAD1

UniProt

Q99259

Additional GAD1/GAD67 Products

Product Documents for GAD1/GAD67 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for GAD1/GAD67 Antibody

For research use only

Citations for GAD1/GAD67 Antibody

Customer Reviews for GAD1/GAD67 Antibody (1)

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  • Human/Mouse/Rat GAD1/GAD67 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Adult brain (hippocampus)
    Species: Mouse
    Verified Customer | Posted 04/28/2023
    We reached out to the customer offering to troubleshoot with no response. Does not work with incubation at 1:500 to 1:1000 dilutions overnight plus secondary AlexaFluor (multiple tested). No fluorescence detected. Sample tissue fixed with PFA.
    GAD1/GAD67 Antibody AF2086
    Bio-Techne Response
    Thank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.

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Protocols

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