ICC/IF for Anti-ROBO1 [IPI-ROBO1.81]
Addgene #251704
- 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 March 13, 2026)
- DOI
- https://doi.org/10.57733/addgene.89j4uw
- Abstract
- Experimental results for Anti-ROBO1 [IPI-ROBO1.81] (Addgene #251704) in an immunofluorescence (IF) assay with COS cells expressing human ROBO1 or ROBO2.
- Citation
-
ICC/IF for Anti-ROBO1 [IPI-ROBO1.81]. Saintpierre A, Nawabi H 2026. Addgene Report, https://doi.org/10.57733/addgene.89j4uw
Image attribution: Institute for Protein Innovation (IPI)
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-ROBO1 [IPI-ROBO1.81]
Addgene #251704 -
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-ROBO1 [IPI-ROBO1.81]
- Source
- Addgene
- Catalog Number
- 251704
- Host Species
- Rabbit
- Target Antigen
- ROBO1
- 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 ROBO1 labeling. Negative for ROBO2 labeling.