Anti-CALB2 [BCN4.2.3B2] in Immunocytochemistry (Transfected COS)
Addgene #204683
- Data Submitted By
- James Trimmer
- Lab Name
- James Trimmer, UC Davis
- Submission Date
- October 10, 2024
- Publication Date
- November 21, 2024 (modified November 21, 2024)
- Abstract
- COS cells were transfected with human Flag-CALB2, fixed and labeled with Anti-Flag (Sigma, #F7425-.2mg), Anti-CALB2 [BCN4.2.3B2] recombinant monoclonal antibody (Addgene #204683), or hybridoma-derived Anti-CALB2 [L22/6] monoclonal antibody. Similar labeling was observed between the [L22/6] monoclonal antibody and the [BCN4.2.3B2] recombinant antibody. The labeling pattern of Flag-CALB2 was confirmed with Anti-Flag antibody.
![COS cells were transfected with human Flag-CALB2, fixed and labeled with Anti-Flag, Anti-CALB2 [BCN4.2.3B2] recombinant monoclonal antibody (R-mAb), or Anti-CALB2 [L22/6] monoclonal antibody (mAb). Nuclei were counterstained with Hoechst.](https://datahub-media.addgene.org/report-images/68xnqc/CALB2.jpg)
Image attribution: James Trimmer, UC Davis
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-CALB2 [BCN4.2.3B2]
Addgene #204683 -
Goat anti-mouse IgG2a cross-adsorbed secondary antibody
Thermo Fisher #A21131
Materials & Methods
Sample
- Species
- Human
- Cell / Tissue Type
- COS
- Additional Information
- COS cells were transfected with human Flag-CALB2
Buffer
- Name
- PBS
- pH
- 7.4
Fixative
- Fixative
- Fixed and permeabilized with 3% paraformaldehyde powder and 0.1% Triton X-100 in PBS/Ca/Mg
- Time
- 30 min
- Temperature
- 4 ˚C
Blocking
- Agent
- Milk
- Concentration
- 4% w/v
- Time
- 45 min
- Temperature
- Room Temperature
Primary Antibody
- Name
- Anti-CALB2 [BCN4.2.3B2]
- Source
- Addgene
- Catalog Number
- 204683
- Species
- Mouse
- Gene / Target
- CALB2
- Concentration
- 5 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 (Link opens in a new window)
- Controls
- Labeling by the recombinant monoclonal antibody was compared to a hybridoma-derived monoclonal antibody against the same target. Anti-Flag labeling (Sigma, #F7425-.2mg) 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 monoclonal antibody and recombinant antibody.