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Anti-GAD1 [BCN49.2.3D1] in Immunocytochemistry (Transfected COS)

Addgene #210068

Data Submitted By: James Trimmer
Lab Name: James Trimmer, UC Davis
Submission Date: June 30, 2024
Publication Date: July 30, 2024 (modified August 21, 2024)
Abstract: COS cells were transfected with GFP-tagged, human GAD1, fixed and labeled with Anti-GAD1 [BCN49.2.3D1] (Addgene #210068) or NeuroMab L127/8. Similar labeling was observed between antibodies and colocalized with GFP.

COS cells were transfected with GFP-tagged, human GAD1, fixed and labeled with Anti-GAD1 [BCN49.2.3D1] (GAD1 BCN49.2.3D1 R-mAb) or NeuroMab L127/8 (L28/8 mAb). Nuclei were counterstained with Hoechst. — Caption: COS cells were transfected with GFP-tagged, human GAD1, fixed and labeled with Anti-GAD1 [BCN49.2.3D1] (GAD1 BCN49.2.3D1 R-mAb) or NeuroMab L127/8 (L28/8 mAb). Nuclei were counterstained with Hoechst.
Image attribution: James Trimmer, UC Davis

This report is made available under the Creative Commons Attribution 4.0 International License.

Antibodies Used: Anti-GAD1 [BCN49.2.3D1]
Addgene #210068; Goat anti-Mouse IgG2a Cross-Adsorbed Secondary Antibody
Thermo Fisher #A21131

Materials & Methods

Sample

Species: Primate
Cell / Tissue Type: COS
Additional Information: COS cells were transfected with GFP-tagged, human GAD1

Buffer

Name: PBS
pH: 7.4

Fixative

Fixative: Formaldehyde/Paraformaldehyde
Time: 30 min
Temperature: 4 ˚C

Blocking

Agent: Milk
Concentration: 4% w/v
Time: 45 min
Temperature: Room Temperature

Primary Antibody

Name: Anti-GAD1 [BCN49.2.3D1]
Source: Addgene
Catalog Number: 210068
Species: Mouse
Concentration: 10 µg/mL
Time: 45 min
Temperature: Room Temperature

View in Addgene Catalog

Secondary Antibody

Name: Goat anti-Mouse IgG2a Cross-Adsorbed Secondary Antibody
Source: Thermo Fisher
Catalog Number: A21131
Concentration: 1:2500
Conjugate: Alexa Fluor 488
Time: 45 min
Temperature: Room Temperature

Additional Information

Protocol: Trimmer lab immunocytochemistry protocol
Controls: Labeling of Anti-GAD1 [BCN49.2.3D1] was compared to the previously validated NeuroMab L127/8. GFP colocalization confirmed the correct protein was labeled.

Results

Result: Pass (The antibody worked under these conditions)
Describe Results: Equivalent signal strength and labeling pattern between antibodies and colocalization with GFP