ICC/IF for Anti-Neuroligin-1 [IPI-NLGN1.31] in binding assay
Addgene #250792
- Data Submitted By
- Alexander Morano , Travis Riedel and Deborah Moshinsky
- Lab Name
- Institute for Protein Innovation (IPI)
- Submission Date
- February 03, 2026
- Publication Date
- February 09, 2026 (modified February 11, 2026)
- Abstract
- Experimental results for Anti-Neuroligin-1 [IPI-NLGN1.31] (Addgene #250792) in an immunofluorescence (IF) binding assay with ExpiCHO cells expressing human or mouse NLGN1.
![A) Immunofluorescence (IF) of ExpiCHO cells transfected with human (top) and mouse (bottom) NLGN1 and labeled with Anti-Neuroligin-1 [IPI-NLGN1.31] (Addgene #250792). Left images show all three channels (IPI-NLGN1.31, GFP (transfection control) and DAPI) while the right images only show IPI-NLGN1.31 and DAPI. Images taken at 40X magnification. B) Combined quantification of multiple images of the same transfected cells taken at 10X magnification. IPI-NLGN1.31 labeling of human (left) and mouse (right) Anti-Neuroligin-1 [IPI-NLGN1.31] is shown in the top row, and compared to a positive (left) and negative (right) control in the bottom row. For both panels, IPI-NLGN1.31 was used at a 1 µg/mL (1:1,000 dilution).](https://datahub-media.addgene.org/report-images/skyikn/NLGN1.31_IF-BINDING.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-Neuroligin-1 [IPI-NLGN1.31]
Addgene #250792 -
Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment
Jackson ImmunoResearch #111-606-047
Materials & Methods
Sample
- Target Species
- Human
- Cell / Tissue Type
- ExpiCHO
- Additional Information
- ExpiCHO cells were transiently transfected with GFP (transfection control) and V5-tagged human NLGN1
Buffer
- Name
- PBS
Fixative
- Fixative
- Formaldehyde/Paraformaldehyde
- Time
- 15 min
- Temperature
- Room Temperature
Permeabilization
- Agent
- Triton X-100 in PBS
- Concentration
- 0.20%
- 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-Neuroligin-1 [IPI-NLGN1.31]
- Source
- Addgene
- Catalog Number
- 250792
- Host Species
- Rabbit
- Target Antigen
- NLGN1
- Concentration
- 1 µg/mL (1:1,000 dilution)
- Time
- 60 min
- Temperature
- Room Temperature
Secondary Antibody
- Name
- Alexa Fluor 647 F(ab')2 goat anti-rabbit IgG Fc fragment
- Source
- Jackson ImmunoResearch
- Catalog Number
- 111-606-047
- Concentration
- 2 µg/mL
- Time
- 60 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
Confocal images taken at 40X magnification on an EV0S-m7000 microscope and deconvolved using the richardson-lucy algorithm in the FIJI deconlab plugin.
For quantitative data, GFP-positive cells were identified via the neural network CellPose, then the mean fluorescence intensity (MFI) of the far-red channel for each cell, representing IPI-NLGN1.31 labeling, was recorded. Each histogram displays the number of cells with MFIs ranging from below 100 (background fluorescence) to 4095 (saturation).
Controls
Positive control: Anti-V5 [IPI-SV5-Pk1] (Addgene #218107) used to confirm IF labeling efficacy (since construct expressing target also has a V5 tag).
Negative control: Rabbit IgG used to confirm transfected construct does not interact /produce signal with every antibody.
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- The antibody shows binding to human and mouse NLGN1.