IF for Anti-Glypican 5 (GPC5) [IPI-GPC5.13]

Addgene #241000

Data Submitted By: Taciani de Almeida Magalhaes and Adrian Salic
Lab Name: Adrian Salic, Harvard Medical School
Submission Date: August 05, 2025
Publication Date: October 21, 2025 (modified October 21, 2025)
Abstract: Experimental results for Anti-Glypican 5 (GPC5) [IPI-GPC5.13] (Addgene #241000) in an immunofluorescence (IF) assay with HEK293T GPC(1–6)KO cells expressing human GPC1, 2, 3, 4, 5 or 6.

Representative epifluorescence images of labeling of HEK293T GPC(1–6)KO cells transfected with fusion of eGFP with either human GPC1, 2, 3, 4, 5, or 6 using Anti-Glypican 5 (GPC5) [IPI-GPC5.13] (Addgene #241000). Anti-Glypican 5 (GPC5) [IPI-GPC5.13] recognizes human GPC5, but not human GPC1, 2, 3, 4, and 6. eGFP (transfection control) is shown in green, Anti-Glypican 5 (GPC5) [IPI-GPC5.13] in magenta, and DAPI (nuclear staining) in blue. — Caption: Representative epifluorescence images of labeling of HEK293T GPC(1–6)KO cells transfected with fusion of eGFP with either human GPC1, 2, 3, 4, 5, or 6 using Anti-Glypican 5 (GPC5) [IPI-GPC5.13] (Addgene #241000). Anti-Glypican 5 (GPC5) [IPI-GPC5.13] recognizes human GPC5, but not human GPC1, 2, 3, 4, and 6. eGFP (transfection control) is shown in green, Anti-Glypican 5 (GPC5) [IPI-GPC5.13] in magenta, and DAPI (nuclear staining) in blue.
Image attribution: Adrian Salic

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; Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568
Thermo Fisher #A-11011

Materials & Methods

Sample

Target Species: Human
Cell / Tissue Type: HEK293T
Additional Information: GPC(1–6)KO cells. Transfected with fusion of eGFP with either human GPC1, 2, 3, 4, 5, or 6.

Buffer

Name: TBS

Fixative

Fixative: Formaldehyde/Paraformaldehyde
Time: 30 min

Permeabilization

Agent: TBS with 0.2% Triton X-100 (TBST)

Primary Antibody

Name: Anti-Glypican 5 (GPC5) [IPI-GPC5.13]
Source: Addgene
Catalog Number: 241000
Host Species: Rabbit
Target Antigen: GPC5
Concentration: 1:500 from a stock solution of 1 mg/mL
Time: 90 min
Temperature: Room Temperature

View in Addgene Catalog

Secondary Antibody

Name: Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568
Source: Thermo Fisher
Catalog Number: A-11011
Host Species: Goat
Concentration: 1:1000
Time: 90 min
Temperature: Room Temperature

Additional Information

Additional Information: Fixation in 3.7% PFA (in PBS) for 30 min; Permeabilization in TBS+BSA (25 mg/mL) with 0.2% Triton X-100; Antibodies were diluted in TBS+BSA (25 mg/mL) with 0.1% Triton X-100
Controls: eGFP-GPC (transfection), DAPI (nuclear)

Results

Result: Pass (The antibody worked under these conditions)
Describe Results: Strong signal for GPC5 detected. No signal for GPC1, 2, 3, 4, or 6.
Knockout Validation: Yes