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Anti-Neurexin-2-alpha [IPI-mNRXN2a.7] in Immunohistochemistry (Mouse)

Addgene #244068

Data Submitted By: Alexander Morano , Travis Riedel and Deborah Moshinsky
Lab Name: Institute for Protein Innovation (IPI)
Submission Date: January 30, 2026
Publication Date: February 09, 2026 (modified February 19, 2026)
DOI: https://doi.org/10.57733/addgene.mzuzje
Abstract: Experimental results for Anti-Neurexin-2-alpha [IPI-mNRXN2a.7] (Addgene #244068) in an immunohistochemistry assay with a mouse brain sample.
Citation: Anti-Neurexin-2-alpha [IPI-mNRXN2a.7] in Immunohistochemistry (Mouse). Morano A, Riedel T, Moshinsky D 2026. Addgene Report, https://doi.org/10.57733/addgene.mzuzje

**Caption:** Immunohistochemistry (IHC) of a 20 micron cryosection of the cerebellum section of an adult mouse brain. Widefield images taken at 4X magnification on an EV0S-m7000 microscope. Left image shows Anti-Neurexin-2-alpha [IPI-mNRXN2a.7] (Addgene #244068) and DAPI while the right image only shows IPI-mNRXN2a.7. Antibody was used at 10 µg/mL (1:100 dilution).
**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-alpha [IPI-mNRXN2a.7]
Addgene #244068; Goat anti-rabbit alexa fluor 647
ThermoFisher #A21245

Materials & Methods

Sample

Target Species: Mouse
Cell / Tissue Type: Brain
Additional Information: 20 µm section

Buffer

Name: PBS

Fixative

Fixative: Paraformaldehyde, 4% in PBS
Time: 1.5 h post-fixation with PFA after perfusion and harvesting
Temperature: 4 ˚C

Permeabilization / Delipidation

Agent: PBS with 0.1% Triton-X 100
Time: 15 min (3x 5 min washes)
Temperature: Room Temperature

Blocking

Agent: Normal Goat Serum
Concentration: 5% in PBST
Time: 1 h
Temperature: Room Temperature

Primary Antibody

Name: Anti-Neurexin-2-alpha [IPI-mNRXN2a.7]
Source: Addgene
Catalog Number: 244068
Host Species: Rabbit
Target Antigen: NRXN2a
Concentration: 10 µg/mL (1:100 dilution)
Time: 60 min
Temperature: Room Temperature

View in Addgene Catalog

Secondary Antibody

Name: Goat anti-rabbit alexa fluor 647
Source: ThermoFisher
Catalog Number: A21245
Concentration: 1:500 dilution from a 2 mg/mL stock
Time: 90 min
Temperature: Room Temperature

Additional Information

Additional Information

Immunohistochemistry Methods: Mouse brain tissue

Adult (3-5 week old) mouse brains were first purchased from Jackson Labs. Instructions were provided to Jackson labs for harvesting and fixing tissue. Anesthesia was performed with inhalant isoflurane, followed by transcardial perfusion with ~11 mL 1X PBS perfusate/mouse containing heparin (10,000 units heparin per 1L 1X PBS), then ~25 mL ice-cold 4% paraformaldehyde in 1X PBS. After dissecting the brain, it was stored in 1X PBS + 0.01% sodium azide solution (NaN3) at 4 °C. Upon arrival of tissue, embryos were washed 3x 10 min in 1X PBS, then cryoprotected overnight in 30% sucrose in 1X PBS, at 4 °C. Adult brains were placed in a 5 mL conical tube containing ice-cold (4 °C) 10% sucrose solution and incubated at 4 °C overnight. The next day, 10% sucrose was replaced with ice-cold 30% sucrose and the brain was allowed to equilibrate completely at 4 °C, as evidenced by its sinking to the bottom of the tube, for 2–3 days.

Equilibrated tissue was then frozen in OCT on dry ice and 20–25 μm sections were cut on a Leica cryostat at the Harvard histology/sectioning core. Slides were thawed for 5 min at RT; sections were rehydrated 1x 5min in 1X PBS and then washed 3x 5 min in 1X PBST (0.1% Triton X-100 in 1X PBS) in a humidified chamber in a slide box. Slides were blocked for 1 h at RT in 5% NGS (Normal Goat Serum) or 5% NDS (Normal Donkey Serum) in 0.1% Triton-X-100 in PBS. Primary antibodies were then diluted in blocking solution and incubated for overnight at 4 °C. Slides were then washed 4x 5min in 0.1% TritonX-100 and incubated with secondary antibodies diluted in blocking solution (5% NGS or NDS) at RT for 1.5–2 h, in the dark. Slides were washed 4x 5 min in 0.1% TritonX-100 again, then 1x 5min in 1X PBS. Slides were then mounted in Fluoromount mounting medium with DAPI, outlined with nail polish to secure the coverslip, and stored at 4 °C in the dark until ready to image.

Controls

Positive control: NeuN counterstain (anti-mouse IGG1).

Negative control: No primary control.

Results

Result: Pass (The antibody worked under these conditions)
Describe Results: This antibody shows distinct labeling above the background in the cerebellum of the mouse brain.