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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.
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