ICC for Anti-Netrin-1 [IPI-NTN1.45]
Addgene #259427
- Data Submitted By
- Melissa Pestemalciyan and Timothy Kennedy
- Lab Name
- Timothy Kennedy, McGill University
- Submission Date
- July 27, 2026
- Publication Date
- August 14, 2026 (modified August 14, 2026)
- DOI
- https://doi.org/10.57733/addgene.idamz9
- Abstract
- Experimental results for Anti-Netrin-1 [IPI-NTN1.45] (Addgene #241895) in an immunofluorescence (IF) binding assay with HEK293 cells expressing chick NTN1.
- Citation
-
ICC for Anti-Netrin-1 [IPI-NTN1.45]. Pestemalciyan M, Kennedy T 2026. Addgene Report, https://doi.org/10.57733/addgene.idamz9
![Immunocytochemistry (ICC) analysis of HEK293 cells expressing chick Netrin-1. Labeling for NTN1 was performed using Anti-Netrin-1 [IPI-NTN1.45] (Addgene #259427). Positive control was an identical sample processed identically in parallel with a commercially available NTN1 antibody (Abcam, #ab126729).](https://datahub-media.addgene.org/report-images/idamz9/NTN1.45_Kennedy_ICC.png)
Image attribution: Timothy Kennedy, McGill University
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-Netrin-1 [IPI-NTN1.45]
Addgene #259427 -
Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 555
Thermo Fisher #A31572
Materials & Methods
Sample
- Target Species
- Chicken
- Cell / Tissue Type
- HEK293 cells
- Additional Information
- HEK293 cell line expressing chick Netrin-1
Buffer
- Name
- PBS
Fixative
- Fixative
- 4% PFA in PBS
- Time
- 15 min
- Temperature
- On ice
Permeabilization
- Agent
- Triton X-100
- Concentration
- 0.25%
- Time
- 15 min
- Temperature
- Room Temperature
Blocking
- Agent
- 5% Heat inactivated horse serum, 0.25% Triton X-100 in PBS
- Time
- 60 min
- Temperature
- Room Temperature
Primary Antibody
- Name
- Anti-Netrin-1 [IPI-NTN1.45]
- Source
- Addgene
- Catalog Number
- 259427
- Host Species
- Rabbit
- Target Antigen
- NTN1
- Concentration
- 2 µg/mL
- Time
- Overnight
- Temperature
- 4 ˚C
Secondary Antibody
- Name
- Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 555
- Source
- Thermo Fisher
- Catalog Number
- A31572
- Concentration
- 0.736 µg/mL
- Conjugate
- AF 555
- Time
- 60 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
Imaged on a Zeiss Axiovert S100TVEpifluorescence microscope.
Positive control was an identical sample processed identically in parallel with the commercially available Anti-Netrin 1 antibody [EPR5428] (Abcam, #ab126729), which is also a rabbit monoclonal antibody. This antibody has been KO validated by Kennedy lab (Bin et al., 2015). The same secondary antibody was used for the positive control.
Negative control (not shown) was performed using only the secondary antibody.
References
Bin et al. (2015). Complete Loss of Netrin-1 Results in Embryonic Lethality and Severe Axon Guidance Defects without Increased Neural Cell Death Cell Rep, 12(7):1099-106. https://doi.org/10.1016/j.celrep.2015.07.028 (Link opens in a new window) PMID: 26257176 (Link opens in a new window)
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- Positive labeling for NTN1, consistent with results from a previously validated antibody.