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Anti-ROBO3 [IPI-ROBO3.119] in Surface Plasmon Resonance (SPR)

Addgene #251720

Data Submitted By
Murali Anuganti , Aditi Kachare , Travis Riedel and Deborah Moshinsky
Lab Name
Institute for Protein Innovation (IPI)
Submission Date
February 06, 2026
Publication Date
March 11, 2026 (modified March 13, 2026)
DOI
https://doi.org/10.57733/addgene.ea1t2g
Abstract
Experimental results for Anti-ROBO3 [IPI-ROBO3.119] (Addgene #251720) in SPR assay with a human sample.
Citation
Anti-ROBO3 [IPI-ROBO3.119] in Surface Plasmon Resonance (SPR). Anuganti M, Kachare A, Riedel T, Moshinsky D 2026. Addgene Report, https://doi.org/10.57733/addgene.ea1t2g
Surface Plasmon Resonance (SPR) kinetics analysis of the interaction between Anti-ROBO3 [IPI-ROBO3.119] and human ROBO3. SPR binding kinetics were measured on a Carterra LSA using HC30M chips (Carterra, #4279) at 25 °C. Goat anti-rabbit IgG Fc (Jackson ImmunoResearch, #111-005-046) was immobilized via amine coupling, and test antibodies were captured using a 96-channel print-head. Antigens (400 nM to five lower concentrations, 2-fold dilutions) were injected in antigen buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 1 mM CaCl₂, 1 mM MgCl₂, 0.005% Tween 80) with 300 s association/ dissociation phases and acid regeneration. Data (reference/buffer subtracted, smoothed) were globally fit to a 1:1 Langmuir model to derive ka, kd, and KD using Carterra Kinetics software v1.9.2.44.63, and replotted in OriginPro 2023b. Results show a high-affinity and specific binding event between the antibody and antigen.
Caption: Surface Plasmon Resonance (SPR) kinetics analysis of the interaction between Anti-ROBO3 [IPI-ROBO3.119] and human ROBO3. SPR binding kinetics were measured on a Carterra LSA using HC30M chips (Carterra, #4279) at 25 °C. Goat anti-rabbit IgG Fc (Jackson ImmunoResearch, #111-005-046) was immobilized via amine coupling, and test antibodies were captured using a 96-channel print-head. Antigens (400 nM to five lower concentrations, 2-fold dilutions) were injected in antigen buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 1 mM CaCl₂, 1 mM MgCl₂, 0.005% Tween 80) with 300 s association/ dissociation phases and acid regeneration. Data (reference/buffer subtracted, smoothed) were globally fit to a 1:1 Langmuir model to derive ka, kd, and KD using Carterra Kinetics software v1.9.2.44.63, and replotted in OriginPro 2023b. Results show a high-affinity and specific binding event between the antibody and antigen.
Image attribution: Institute for Protein Innovation (IPI)

This report is made available under the Creative Commons Attribution 4.0 International License.

Antibody Used
Anti-ROBO3 [IPI-ROBO3.119]
Addgene #251720

Materials & Methods

Sample

Target Species
Human

Primary Antibody

Name
Anti-ROBO3 [IPI-ROBO3.119]
Source
Addgene
Catalog Number
251720
Host Species
Rabbit
Target Antigen
ROBO3
Concentration
0.25 µg/mL (1:4,000 dilution)
Temperature
Room Temperature
View in Addgene Catalog

Results

Result
Pass (The antibody worked under these conditions)
Describe Results
Results show a high-affinity and specific binding event between the antibody and antigen.