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Anti-Versican [N351/23R] in Immunocytochemistry (Transfected COS)

Addgene #209585

Data Submitted By
James Trimmer
Lab Name
James Trimmer, UC Davis
Submission Date
October 10, 2024
Publication Date
October 18, 2024 (modified November 22, 2024)
Abstract
COS cells were transfected with mouse versican, fixed and labeled with Anti-Versican [N351/23R] recombinant monoclonal antibody (Addgene #209585) or its hybridoma-derived monoclonal antibody parent. Similar labeling was observed between the recombinant antibody and its monoclonal antibody parent.
Caption: COS cells were transfected with mouse versican, fixed and labeled with Anti-Versican [N351/23R] recombinant monoclonal antibody (R-mAb) or its hybridoma-derived monoclonal antibody parent (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-Versican [N351/23R]
Addgene #209585
Goat anti-mouse IgG2a cross-adsorbed secondary antibody
Thermo Fisher #A21131

Materials & Methods

Sample

Species
Mouse
Cell / Tissue Type
COS
Additional Information
COS cells were transfected with mouse versican

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-Versican [N351/23R]
Source
Addgene
Catalog Number
209585
Species
Mouse
Gene / Target
Vcan
Concentration
4 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 (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.

Results

Result
Pass (The antibody worked under these conditions)
Describe Results
Equivalent signal strength and labeling pattern between parental monoclonal antibody and recombinant antibody.