Anti-Progranulin antibody [EPR15864]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progranulin antibody [EPR15864] (AB208777)

Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labeling Progranulin with ab208777 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human gastric cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 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-Progranulin antibody [EPR15864] (AB208777)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Progranulin with ab208777 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Kupffer cells of Human liver is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 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-Progranulin antibody [EPR15864] (AB208777)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Progranulin with ab208777 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on trophoblastic cells of Human placenta is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 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-Progranulin antibody [EPR15864] (AB208777)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Progranulin with ab208777 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human breast cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab97051 at 1/500 dilution.

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

• IP

Supplier Data

Immunoprecipitation - Anti-Progranulin antibody [EPR15864] (AB208777)

Progranulin was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab208777 at 1/70 dilution.

Western blot was performed from the immunoprecipitate using ab208777 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

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

Lane 2 : ab208777 IP in HeLa whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal -Isotype Control (ab172730) instead of ab208777 in HeLa whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-Progranulin antibody [EPR15864] (ab208777)

Predicted band size: 64 kDa

false

• WB

Supplier Data

Western blot - Anti-Granulin antibody [EPR15864] (AB208777)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lane 1:

Western blot - Anti-Granulin antibody [EPR15864] (ab208777) at 1/2000 dilution

Lane 2:

Western blot - Anti-Granulin antibody [EPR15864] (ab208777) at 1/10000 dilution

Lane 1:

A431 (Human epidermoid carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 64 kDa

Observed band size: 74 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-Progranulin antibody [EPR15864] (AB208777)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Progranulin antibody [EPR15864] (ab208777) at 1/2000 dilution

All lanes:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 64 kDa

Observed band size: 74 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Progranulin antibody [EPR15864] (AB208777)

Lanes 1 - 3 : Merged signal (red and green). Green - ab208777 observed at 64 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab208777 was shown to specifically react with Progranulin in wild-type HAP1 cells as signal was lost in GRN (Progranulin) knockout cells. Wild-type and GRN (Granulin) knockout samples were subjected to SDS-PAGE. ab208777 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Progranulin antibody [EPR15864] (ab208777) at 1/2000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

GRN (Granulin) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Predicted band size: 64 kDa

false

• WB

Lab

Western blot - Anti-Progranulin antibody [EPR15864] (AB208777)

Lanes 1-4 : Merged signal (red and green). Green - ab208777 observed at 74 kDa. Red - loading control ab8245 observed at 36 kDa.

ab208777 Anti-Progranulin antibody [EPR15864] was shown to specifically react with Granulin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266738 (knockout cell lysate ab257235) was used. Wild-type and Progranulin knockout samples were subjected to SDS-PAGE. ab208777 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-Progranulin antibody [EPR15864] (ab208777) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

GRN knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-grn-granulin-knockout-hek-293t-cell-line-ab266738'>ab266738</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HAP1 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: 64 kDa

Observed band size: 74 kDa

false

• WB

Lab

Western blot - Anti-Progranulin antibody [EPR15864] (AB208777)

ab208777 was shown to react with GRN in wild-type HEK293T cells in Western blot with loss of signal observed in GRN knockout cell line ab266738. Wild-type HEK293T and GRN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab208777 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Progranulin antibody [EPR15864] (ab208777) at 1/1000 dilution

Lane 1:

Wild-type HEK293T lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-grn-granulin-knockout-hek-293t-cell-line-ab266738'>ab266738</a>)

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-grn-granulin-knockout-hek-293t-cell-lysate-ab257235'>ab257235</a>)

Observed band size: 64 kDa

false

• WB

Supplier Data

Western blot - Anti-Progranulin antibody [EPR15864] (AB208777)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Progranulin antibody [EPR15864] (ab208777) at 1/1000 dilution

All lanes:

Human GRN full-length recombinant protein with GST-tag at N-terminal at 0.01 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 64 kDa

Observed band size: 90 kDa

false

Exposure time: 15s