PA5-47256

antibody from Invitrogen Antibodies

Targeting: GPC3 DGSX, OCI-5, SDYS, SGB, SGBS, SGBS1

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Recommended by provider)

Antibody Data

Product number

PA5-47256

Provider

Invitrogen Antibodies

Product name

Glypican 3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs, less than 10% cross-reactivity with recombinant mouse Glypican 3 is observed and less than 5% cross-reactivity with recombinant human (rh) Glypican 2, rhGlypican 5, and rhGlypican 6 is observed. Reconstitute at 0.2 mg/mL in sterile PBS.

Reactivity

Human

Host

Sheep

Isotype

IgG

Vial size

100 µg

Concentration

0.2 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Transferrin Receptor Is a Specific Ferroptosis Marker.

Feng H, Schorpp K, Jin J, Yozwiak CE, Hoffstrom BG, Decker AM, Rajbhandari P, Stokes ME, Bender HG, Csuka JM, Upadhyayula PS, Canoll P, Uchida K, Soni RK, Hadian K, Stockwell BR
Cell reports 2020 Mar 10;30(10):3411-3423.e7

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis from lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-human Glypican 3 Antigen Affinity-purified Polyclonal Antibody (Product # PA5-47256) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody. A specific band was detected for Glypican 3 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Glypican 3 in HepG2 human hepatocellular carcinoma cell line. Samples were incubated in Glypican 3 polyclonal antibody (Product # PA5-47256) using a dilution of 2 µg/mL followed by a HRP-conjugated Anti-Sheep IgG secondary antibody. A specific band was detected for Glypican 3 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Glypican 3 Polyclonal Antibody (Product # PA5-47256) and a 66 kDa band corresponding to GPC3 was observed only in Hep G2. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3) and T-47D (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Rabbit anti-Sheep IgG (H+L), Secondary Antibody, HRP (Product # 61-8620, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

ELISA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

ELISA of Glypican 3 in recombinant Glypican 3 protein. Samples were incubated in Glypican 3 polyclonal antibody (Product # PA5-47256). Protein was serially diluted 2-fold. Antibody was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP followed by Substrate Solution and stopping the enzymatic reaction with Stop Solution.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of Glypican 3 in immersion fixed HepG2 human hepatocellular carcinoma cell line. Samples were incubated in Glypican 3 polyclonal antibody (Product # PA5-47256) using a dilution of 1.7 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.