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IF for Anti-Glypican 1 (GPC1) [IPI-GPC1.21]

Addgene #222511

Data Submitted By
Taciani de Almeida Magalhaes and Adrian Salic
Lab Name
Adrian Salic, Harvard Medical School
Submission Date
October 15, 2024
Publication Date
November 06, 2024 (modified November 11, 2024)
DOI
https://doi.org/10.57733/addgene.z29s5s
Abstract
Representative confocal images of anti-GPC1 labeling of HEK293T beta-1,3-glucuronyltransferase 3 (B3GAT3) KO cells transfected with eGFP-SMO (negative control) or eGFP-GPC1 using a human IgG form of the Anti-Glypican 1 (GPC1) [IPI-GPC1.21] (Addgene #222511).
Citation
IF for Anti-Glypican 1 (GPC1) [IPI-GPC1.21]. de Almeida Magalhaes T, Salic A 2024. Addgene Report, https://doi.org/10.57733/addgene.z29s5s
Caption: Representative confocal images of anti-GPC1 labeling of HEK293T beta-1,3-glucuronyltransferase 3 (B3GAT3) KO cells expressing GPC1. Cells were transfected with eGFP-SMO or eGFP-GPC1. A human IgG form of Anti-Glypican 1 (GPC1) [IPI-GPC1.21] (Addgene #222511) was used to label GPC1 (red).
Image attribution: Adrian Salic, PhD

This report is made available under the Creative Commons Attribution 4.0 International License.

Antibodies Used
Anti-Glypican 1 (GPC1) [IPI-GPC1.21]
Addgene #222511
Goat anti-human 568nm
Thermo Fisher #A-21090

Materials & Methods

Sample

Species
Human
Cell / Tissue Type
HEK293T beta-1,3-glucuronyltransferase 3 (B3GAT3) KO cells
Additional Information
B3GAT3 is required for attachment of both heparin sulfate (HS) and chondroitin sulfate (CS) chains to the GPC protein core

Buffer

Name
TBS

Fixative

Fixative
Formaldehyde/Paraformaldehyde
Time
30 min

Permeabilization

Agent
TBS+BSA (25 mg/mL) with 0.2% Triton X-100

Primary Antibody

Name
Anti-Glypican 1 (GPC1) [IPI-GPC1.21]
Source
Addgene
Catalog Number
222511
Gene / Target
GPC1
Concentration
1:500
Time
90 min
Temperature
Room Temperature
View in Addgene Catalog

Secondary Antibody

Name
Goat anti-human 568nm
Source
Thermo Fisher
Catalog Number
A-21090
Species
Goat
Concentration
1:1000
Time
90 min
Temperature
Room Temperature

Additional Information

Additional Information

Fixation in 3.7% PFA (in PBS); 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 (positive), DAPI (nuclear), eGFP-SMO (negative)

Results

Result
Pass (The antibody worked under these conditions)
Describe Results

NOTE: This data was collected using a human IgG isotype of Anti-Glypican 1 (GPC1) [IPI-GPC1.21], which was found to perform equally to the available rabbit IgG version (Addgene #222511).

Strong signal for target.