ICC/IF for Anti-SEMA3A [IPI-SEM3A.17]
Addgene #254646
- Data Submitted By
- Goenuel Seyit-Bremer and Ruediger Klein
- Lab Name
- Ruediger Klein, Max Planck Institute for Biological Intelligence
- Submission Date
- April 16, 2026
- Publication Date
- May 12, 2026 (modified May 14, 2026)
- DOI
- https://doi.org/10.57733/addgene.qirlhs
- Abstract
- Experimental results for Anti-SEMA3A [IPI-SEM3A.17] (Addgene #254646) in an immunofluorescence (IF) assay with Neuro-2a cells expressing human SEMA3A.
- Citation
-
ICC/IF for Anti-SEMA3A [IPI-SEM3A.17]. Seyit-Bremer G, Klein R 2026. Addgene Report, https://doi.org/10.57733/addgene.qirlhs
![Immunoflourescene for Anti-SEMA3A [IPI-SEM3A.17] (Addgene #254646). Neuro-2a cells were transfected with human SEMA3A-FLAG (SEMA3A + GFP overexpression; top) or with control (GFP overexpression; bottom). Cells were treated with IPI-SEM3A.17 and imaged. Results show strong labeling for cells expressing SEMA3A but none for control cells. Commercial SEMA3A antiobody (Abcam, ab23393) was also tested and resulted in the same labeling patterns (data not shown).](https://datahub-media.addgene.org/report-images/qirlhs/Klein-SEMA3A17.png)
Image attribution: Ruediger Klein, Max Planck Institute for Biological Intelligence
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-SEMA3A [IPI-SEM3A.17]
Addgene #254646 -
Donkey Anti-Rabbit IgG (H+L)
Jackson ImmunoResearch #711-165-152
Materials & Methods
Sample
- Target Species
- Human
- Cell / Tissue Type
- Neuro-2a cells (mouse neuroblasts)
- Additional Information
- Neuro-2a cells were transfected with human SEMA3A-FLAG and GFP.
Buffer
- Name
- PBS
- pH
- 7.4
Fixative
- Fixative
- Formaldehyde/Paraformaldehyde
- Time
- 10 min
- Temperature
- Room Temperature
Permeabilization
- Agent
- Triton X-100 in PBS
- Concentration
- 0.10%
- Time
- 10 min
- Temperature
- Room Temperature
Blocking
- Agent
- BSA
- Concentration
- 1% BSA
- Time
- 30 min
- Temperature
- Room Temperature
Primary Antibody
- Name
- Anti-SEMA3A [IPI-SEM3A.17]
- Source
- Addgene
- Catalog Number
- 254646
- Host Species
- Rabbit
- Target Antigen
- SEMA3A
- Concentration
- 5 µg/mL (1:200 dilution)
- Time
- Overnight
- Temperature
- 4 ˚C
Secondary Antibody
- Name
- Donkey Anti-Rabbit IgG (H+L)
- Source
- Jackson ImmunoResearch
- Catalog Number
- 711-165-152
- Concentration
- 1:200 dilution from 1.5 mg/mL stock
- Conjugate
- Cy3
- Time
- 60 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
Controls
Positive controls: Rabbit Anti-Semaphorin 3A antibody (Abcam, ab23393), and Mouse Anti-FLAG M2 antibody (Sigma, F3165).
Negative control: GFP overexpressing cells.
"
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- Strong signal