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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
Caption: Immunofluorescence (IF) of COS cells transfected with myc-tagged human ROBO1 or ROBO2 and labeled with Anti-ROBO1 [IPI-ROBO1.81] (Addgene #251704).
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
View in Addgene Catalog

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.