ICC/IF for Anti-ROBO2 [IPI-ROBO2.47]
Addgene #251719
- Data Submitted By
- Ana Saintpierre and Homaira Nawabi
- Lab Name
- Homaira Nawabi, University Grenoble-Alpes
- Submission Date
- February 11, 2026
- Publication Date
- March 11, 2026 (modified July 14, 2026)
- DOI
- https://doi.org/10.57733/addgene.b9zkoq
- Abstract
- Experimental results for Anti-ROBO2 [IPI-ROBO2.47] (Addgene #251719) in an immunofluorescence (IF) assay with COS cells expressing human ROBO1 or ROBO2.
- Citation
-
ICC/IF for Anti-ROBO2 [IPI-ROBO2.47]. Saintpierre A, Nawabi H 2026. Addgene Report, https://doi.org/10.57733/addgene.b9zkoq
![Immunofluorescence (IF) of COS cells transfected with myc-tagged human ROBO1 or ROBO2 and labeled with Anti-ROBO2 [IPI-ROBO2.47] (Addgene #251719).](https://datahub-media.addgene.org/report-images/b9zkoq/ROBO2.47_IF_Nawabi.png)
Image attribution: Institute for Protein Innovation (IPI)
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-ROBO2 [IPI-ROBO2.47]
Addgene #251719 -
Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment
Jackson ImmunoResearch #111-606-046
Materials & Methods
Sample
- Target Species
- Human
- Cell / Tissue Type
- COS
- Additional Information
- Cos cells were seeded on coverslips and when 70% of confluence was reached, cells were transfected for 48h with plasmids expressing myc-tagged human ROBO1 or ROBO2 (3 µg of DNA), using PEI transfection reagent.
Buffer
- Name
- PBS
Fixative
- Fixative
- Formaldehyde/Paraformaldehyde
- Time
- 15 min
- Temperature
- Room Temperature
Permeabilization
- Agent
- Triton X-100 in PBS
- Concentration
- 0.10%
- Time
- 10 min
- Temperature
- Room Temperature
Blocking
- Agent
- Normal goat serum + BSA
- Concentration
- 5% serum, 5% BSA
- Time
- 30 min
- Temperature
- Room Temperature
Primary Antibody
- Name
- Anti-ROBO2 [IPI-ROBO2.47]
- Source
- Addgene
- Catalog Number
- 251719
- Host Species
- Rabbit
- Target Antigen
- ROBO2
- Concentration
- 2 µg/mL (1:500 dilution)
- Time
- 120 min
- Temperature
- Room Temperature
Secondary Antibody
- Name
- Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment
- Source
- Jackson ImmunoResearch
- Catalog Number
- 111-606-046
- Concentration
- 2 µg/mL
- Time
- 60 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
Positive control: Anti-myc tag
Negative control: No transfection
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- Positive for ROBO2 labeling. Negative for ROBO1 labeling.