ICC/IF for Anti-Glypican 4 (GPC4) [IPI-mGPC4.3] in human cell lines

Addgene #240999

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 4 (GPC4) [IPI-mGPC4.3] (Addgene #240999) in an immunofluorescence (IF) endogenous assay with human glioblastoma (U-188 MG) and neuroblastoma (SH-SY5Y) cell lines.

Immunofluorescence (IF) of U-118 MG cells (top row) and SH-SY-5Y cells (bottom row) labeled for GPC4 using Anti-Glypican 4 (GPC4) [IPI-mGPC4.3] (Addgene #240999) 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-mGPC4.3, actin, and DAPI) while the right images only show IPI-mGPC4.3 and DAPI. U-188 MG cells are predicted to have high GPC4 expression, whereas SH-SY5Y cells are predicted to have low GPC4 expression, based on publicly available data (DepMap.org). For both panels, IPI-mGPC4.3 was used at 5 μg/mL (1:200 dilution). — Caption: Immunofluorescence (IF) of U-118 MG cells (top row) and SH-SY-5Y cells (bottom row) labeled for GPC4 using Anti-Glypican 4 (GPC4) [IPI-mGPC4.3] (Addgene #240999) 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-mGPC4.3, actin, and DAPI) while the right images only show IPI-mGPC4.3 and DAPI. U-188 MG cells are predicted to have high GPC4 expression, whereas SH-SY5Y cells are predicted to have low GPC4 expression, based on publicly available data (DepMap.org). For both panels, IPI-mGPC4.3 was used at 5 μg/mL (1:200 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-Glypican 4 (GPC4) [IPI-mGPC4.3]
Addgene #240999; 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: U-118 MG, 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 4 (GPC4) [IPI-mGPC4.3]
Source: Addgene
Catalog Number: 240999
Host Species: Rabbit
Target Antigen: GPC4
Concentration: 5 µg/mL
Time: 60 min
Temperature: Room Temperature

View in Addgene Catalog

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 GPC4 expression (U118-MG).