Anti-PEG10/EDR antibody [EPR20051]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Negative staining on normal cells in human liver.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on part of the cells in human liver cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on the germ cells in human testis is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on some cells in human placenta is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PEG10/EDR with ab215035 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

PEG10/EDR was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab215035 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215035 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : HepG2 whole cell lysate, 10 μg (Input).

Lane 2 : ab215035 IP in HepG2 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab215035 in HepG2 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Predicted band size: 80 kDa

false

• WB

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Western blot - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression of PEG10/EDR in HepG2 cells is consistent with what has been described in the literature PMID : 17942406.

Lanes 1 - 2:

Western blot - Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/2000 dilution

Lane 3:

Western blot - Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/10000 dilution

Lane 1:

HEK 293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 10 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: 80 kDa

Observed band size: 100 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Lanes 1-4 : Merged signal (red and green). Green - ab215035 observed at 100 kDa. Red - loading control ab7291 observed at 50 kDa.

ab215035 Anti-PEG10/EDR antibody [EPR20051] was shown to specifically react with PEG10/EDR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265925 (knockout cell lysate ab258103) was used. Wild-type and PEG10/EDR knockout samples were subjected to SDS-PAGE. ab215035 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 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-PEG10/EDR antibody [EPR20051] (ab215035) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PEG10 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PEG10 (EDR) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-peg10-edr-knockout-hela-cell-line-ab265925'>ab265925</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 80 kDa

Observed band size: 100 kDa

false

• WB

Supplier Data

Western blot - Anti-PEG10/EDR antibody [EPR20051] (AB215035)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/1000 dilution

All lanes:

Human testis lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 80 kDa

Observed band size: 100 kDa

false

Exposure time: 1min