IF for Anti-Glypican 3 (GPC3) [IPI-mGPC3.13]
Addgene #222514
- Data Submitted By
- Taciani de Almeida Magalhaes and Adrian Salic
- Lab Name
- Adrian Salic, Harvard Medical School
- Submission Date
- October 25, 2024
- Publication Date
- November 06, 2024 (modified November 11, 2024)
- DOI
- https://doi.org/10.57733/addgene.9krw4c
- Abstract
- Representative confocal images of anti-GPC3 labeling of HEK293T beta-1,3-glucuronyltransferase 3 (B3GAT3) KO cells transfected with eGFP-SMO (negative control) or eGFP-GPC3 using a human IgG form of Anti-Glypican 3 (GPC3) [IPI-mGPC3.13] (Addgene #222514).
- Citation
-
IF for Anti-Glypican 3 (GPC3) [IPI-mGPC3.13]. de Almeida Magalhaes T, Salic A 2024. Addgene Report, https://doi.org/10.57733/addgene.9krw4c
![Representative confocal images of anti-GPC3 labeling of HEK293T beta-1,3-glucuronyltransferase 3 (B3GAT3) KO cells expressing GPC3. Cells were transfected with eGFP-SMO or eGFP-GPC3. A human IgG form of Anti-Glypican 3 (GPC3) [IPI-mGPC3.13] (Addgene #222514) was used to label GPC3 (red).](https://datahub-media.addgene.org/report-images/9krw4c/GPC3.13_IF-Salic.png)
Image attribution: Adrian Salic, PhD
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-Glypican 3 (GPC3) [IPI-mGPC3.13]
Addgene #222514 -
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 3 (GPC3) [IPI-mGPC3.13]
- Source
- Addgene
- Catalog Number
- 222514
- Gene / Target
- GPC3
- Concentration
- 1:500
- Time
- 90 min
- Temperature
- Room Temperature
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 3 (GPC3) [IPI-mGPC3.13], which was found to perform equally to the available rabbit IgG version (Addgene #222514).
Strong signal for target.