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Anti-Slit-1 [IPI-SLIT1.75] in Surface Plasmon Resonance (SPR)

Addgene #259424

Data Submitted By
Murali Anuganti, Aditi Kachare, Travis Riedel and Deborah Moshinsky
Lab Name
Institute for Protein Innovation (IPI)
Submission Date
July 27, 2026
Publication Date
August 05, 2026 (modified August 14, 2026)
DOI
https://doi.org/10.57733/addgene.bybr4q
Abstract
Experimental results for Anti-Slit-1 [IPI-SLIT1.75] (Addgene #259424) in SPR assay with a human sample.
Citation
Anti-Slit-1 [IPI-SLIT1.75] in Surface Plasmon Resonance (SPR). Riedel T, Moshinsky D, Anuganti M, Kachare A 2026. Addgene Report, https://doi.org/10.57733/addgene.bybr4q
Surface Plasmon Resonance (SPR) kinetics analysis of the interaction between Anti-Slit-1 [IPI-SLIT1.75] (Addgene #259424) and human Slit-1. SPR binding kinetics were measured on a Carterra LSA using HC30M chips (Carterra, cat. #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. (Right) IPI-Slit1.75 was originally generated against the hNSlit1 antigen. Cross-reactivity assessment was performed using antibody samples against mSlit3C, hNSlit3, and hSlit2C andantigens. No binding was observed to any of the tested Slit2 or Slit3 constructs, demonstrating that IPI-Slit1.75 is highly specific for the hNSlit1 antigen. As shown in the SPR data below, all tested Slit2 and Slit3 antigens exhibited flat sensorgram responses, indicating a lack of detectable interaction.
Caption: Surface Plasmon Resonance (SPR) kinetics analysis of the interaction between Anti-Slit-1 [IPI-SLIT1.75] (Addgene #259424) and human Slit-1. SPR binding kinetics were measured on a Carterra LSA using HC30M chips (Carterra, cat. #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. (Right) IPI-Slit1.75 was originally generated against the hNSlit1 antigen. Cross-reactivity assessment was performed using antibody samples against mSlit3C, hNSlit3, and hSlit2C andantigens. No binding was observed to any of the tested Slit2 or Slit3 constructs, demonstrating that IPI-Slit1.75 is highly specific for the hNSlit1 antigen. As shown in the SPR data below, all tested Slit2 and Slit3 antigens exhibited flat sensorgram responses, indicating a lack of detectable interaction.
Image attribution: Institute for Protein Innovation (IPI)

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

Antibody Used
Anti-Slit-1 [IPI-SLIT1.75]
Addgene #259424

Materials & Methods

Sample

Target Species
Human

Primary Antibody

Name
Anti-Slit-1 [IPI-SLIT1.75]
Source
Addgene
Catalog Number
259424
Host Species
Rabbit
Target Antigen
Slit-1
Concentration
4 µg/mL (1:250 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. Binding for Slit2 and Slit3 was not observed.