Anti-SynCAM1 [L45/30R] in Immunocytochemistry (Transfected COS)
Addgene #209587
- 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 HA-tagged, mouse SynCAM1, fixed and labeled with Anti-HA, Anti-SynCAM1 [L45/30] monoclonal antibody, or Anti-SynCAM1 [L45/30R] (Addgene #209587) recombinant monoclonal antibody. Similar labeling was observed between the parental monoclonal antibody and its derivative recombinant antibody. The labeling pattern of HA-tagged, mouse SynCAM1 was confirmed with Anti-HA antibody.
Image attribution: James Trimmer, UC Davis
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-SynCAM1 [L45/30R]
Addgene #209587 -
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 HA-tagged, mouse SynCAM1
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-SynCAM1 [L45/30R]
- Source
- Addgene
- Catalog Number
- 209587
- Species
- Mouse
- Concentration
- 1 µg/mL
- Time
- 45 min
- Temperature
- Room Temperature
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 the recombinant monoclonal antibody was compared to the previously validated monoclonal parent. Anti-HA labeling was used to confirm the correct labeling of the transfected protein.
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- Equivalent signal strength and labeling pattern between parental monoclonal antibody and recombinant antibody