ICC/IF for Anti-Neurexin-2-beta [IPI-NRXN2b.9] (Addgene #273809) in siRNA KD assay
Addgene #237809
- Data Submitted By
- Alexander Morano, Travis Riedel and Deborah Moshinsky
- Lab Name
- Institute for Protein Innovation (IPI)
- Submission Date
- November 12, 2025
- Publication Date
- December 11, 2025 (modified December 11, 2025)
- DOI
- https://doi.org/10.57733/addgene.gf6zps
- Abstract
- Experimental results for Neurexin-2-beta [IPI-NRXN2b.9] (Addgene #273809) in an immunofluorescence (IF) assay with NCIH446 cells treated with siRNA against NRXN2.
- Citation
-
ICC/IF for Anti-Neurexin-2-beta [IPI-NRXN2b.9] (Addgene #273809) in siRNA KD assay. Riedel T, Moshinsky D, Morano A 2025. Addgene Report, https://doi.org/10.57733/addgene.gf6zps
![(A) Immunofluorescence (IF) of NCIH446 cells treated with 20 nM siRNA against the housekeeping gene cyclophilin B (20 nM si-CYCB) or 20 nM siRNA against NRXN2 (20 nM si-NRXN2) for 90 hours, then probed for NRXN2 expression using Anti-Neurexin-2-beta [IPI-NRXN2b.9] (Addgene #237809) and for actin with Alexa Fluor 488-conjugated phalloidin. (B) Immunofluorescence (IF) of NCIH446 cells treated with 20 nM scrambled siRNA (20 nM siSCR) or 20 nM siRNA against housekeeping gene cyclophilin B (20 nM si-CYCB) for 90 hours, then probed for cyclophilin B expression using a commercial α-Cyclophilin B antibody and for actin with Alexa Fluor 488-conjugated phalloidin. Confocal images taken at 40X magnification on an ImageXpress HT confocal.ai microscope. Left images show all 3 channels (IPI-NRXN2b.9 or α-Cyclophilin B, actin, and DAPI) while the right images only show IPI-NRXN2b.9 or α-Cyclophilin B and DAPI. For panel A, NRXN2b.9 was used at 5 µg/mL (1:200 dilution).](https://datahub-media.addgene.org/report-images/gf6zps/NRXN2b.9_IPI-IF-KD_v2.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-Neurexin-2-beta [IPI-NRXN2b.9]
Addgene #237809 -
Alexa Fluor 647 F(ab')2 fragment goat anti-rabbit IgG, Fc fragment-specific
Jackson ImmunoResearch #111-606-046
Materials & Methods
Sample
- Target Species
- Mouse
- Cell / Tissue Type
- NCIH446 cells
- Additional Information
- NCIH446 cells were treated with siRNA against housekeeping gene cyclophilin B (Horizon Research #D-001820-10-05) or against NRXN2 (Horizon Research #L-012905-01-0005).
Buffer
- Name
- PBS
Fixative
- Fixative
- Formaldehyde/Paraformaldehyde
- Time
- 15 min
- Temperature
- Room Temperature
Permeabilization
- Agent
- Triton X-100
- 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-Neurexin-2-beta [IPI-NRXN2b.9]
- Source
- Addgene
- Catalog Number
- 237809
- Host Species
- Rabbit
- Target Antigen
- NRXN2b
- Concentration
- 5 µg/mL
- Time
- 60 min
- Temperature
- Room Temperature
Secondary Antibody
- Name
- Alexa Fluor 647 F(ab')2 fragment goat anti-rabbit IgG, Fc fragment-specific
- Source
- Jackson ImmunoResearch
- Catalog Number
- 111-606-046
- Host Species
- Goat
- Concentration
- 2 µg/mL
- Conjugate
- Alexa Fluor 647
- Time
- 60 min
- Temperature
- Room Temperature
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- siRNA knockdown resulted in reduced labeling with IPI-NRXN2b.9 by a significant degree in cells expressing NRXN2 using a siRNA specific to NRXN2 but not with control siRNA.
- Knockout Validation
- Yes