ICC/IF for Anti-Glypican 5 (GPC5) [IPI-GPC5.13] in human cell lines
Addgene #241000
- Data Submitted By
- Alexander Morano , Travis Riedel and Deborah Moshinsky
- Lab Name
- Institute for Protein Innovation (IPI)
- Submission Date
- October 03, 2025
- Publication Date
- October 03, 2025 (modified October 22, 2025)
- Abstract
- Experimental results for Anti-Glypican 5 (GPC5) [IPI-GPC5.13] (Addgene #241000) in an immunofluorescence (IF) endogenous assay with human small cell lung carcinoma (DMS 114) and neuroblastoma (SH-SY5Y) cell lines.
![Immunofluorescence (IF) of DMS114 cells (top row) and SH-SY-5Y cells (bottom row) labeled for GPC5 using Anti-Glypican 5 (GPC5) [IPI-GPC5.13] (Addgene #241000) 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-GPC5.13, actin, and DAPI) while the right images only show IPI-GPC5.13 and DAPI. DMS114 cells are predicted to have high GPC5 expression, whereas SH-SY5Y cells are predicted to have low GPC5 expression, based on publicly available data (DepMap.org). For both panels, IPI-GPC5.13 was used at 5 μg/mL (1:200 dilution).](https://datahub-media.addgene.org/report-images/g92gk8/GPC5.13_IF_Endogenous.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-Glypican 5 (GPC5) [IPI-GPC5.13]
Addgene #241000 -
Alexa Fluor 647 F(ab')2 fragment goat anti-rabbit IgG, Fc fragment-specific
Jackson ImmunoResearch #111-606-046
Materials & Methods
Sample
- Target Species
- Human
- Cell / Tissue Type
- DMS 114, SH-SY5Y
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-Glypican 5 (GPC5) [IPI-GPC5.13]
- Source
- Addgene
- Catalog Number
- 241000
- Host Species
- Rabbit
- Target Antigen
- GPC5
- 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
- Concentration
- 2 µg/mL
- Time
- 60 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
Confocal images taken at 40X magnification on an ImageXpress HT confocal.ai microscope. The same imaging parameters and antibody concentrations were used for the labeling and imaging of each cell culture.
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- The antibody shows binding in human cancer cell lines with expected high GPC5 expression (U118-MG).