Anti-Poliovirus Receptor/PVR antibody [EPR22672-151]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Poliovirus Receptor/PVR with ab267788 at 1/500 dilution (1.01μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human colon cancer (PMID : 28816021,11454801). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt

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Flow Cytometry - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Flow cytometric analysis of Raji (Human Burkitt's lymphoma B lymphocyte) (Left panel) and A549 (Human lung carcinoma epithelial cell) (Right panel) labeling Poliovirus Receptor/PVR with ab267788 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : Raji (PMID : 31500665).

Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling Poliovirus Receptor/PVR with ab267788 at 1/500 dilution (1.01μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer (PMID : 20331633). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Poliovirus Receptor/PVR with ab267788 at 1/500 dilution (1.01μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human breast cancer (PMID : 31035013). Counterstained with hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

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Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

PVR/CD155 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 17893876,11454801).

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (ab267788) at 1/1000 dilution

Lane 1:

Human placenta lysate at 20 µg

Lane 2:

Human lung cancer lysate at 20 µg

Lane 3:

Human colon cancer lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/2000 dilution

Predicted band size: 45 kDa

Observed band size: 70 kDa

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• WB

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Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Lanes 1- 4 : Merged signal (red and green). Green - ab267788 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab267788 was shown to react with Poliovirus Receptor/PVR in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266102 (knockout cell lysate ab257622) was used. Wild-type HEK-293T and PVR knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab267788 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (ab267788) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PVR knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PVR (Poliovirus Receptor) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-pvr-poliovirus-receptor-knockout-hek-293t-cell-line-ab266102'>ab266102</a>)

Lane 3:

Wild-type A549 cell lysate at 20 µg

Lane 4:

PVR knockout A549 cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 70 kDa

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• WB

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Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (AB267788)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 10 seconds.

Negative control : Raji (PMID : 28829151, 31500665).

PVR/CD155 is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 17893876,11454801).

All lanes:

Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR22672-151] (ab267788) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

WI-38 (human fetal lung fibroblast), whole cell lysate at 20 µg

Lane 3:

HT-1080 (human fibrosarcoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

HUVEC (human umbilical vein endothelial cell), whole cell lysate at 20 µg

Lane 6:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 7:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 70 kDa

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