Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Granulin 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab208777).

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-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Granulin 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab208777).

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-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

This IHC data was generated using the same anti-Granulin antibody clone [EPR15864] in a different buffer formulation (cat# ab208777).

Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labeling Granulin 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-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Granulin 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab208777).

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

• WB

Lab

Western blot - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

This data was developed using the same antibody clone in a different buffer formulation (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-Granulin 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 Granulin 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-Granulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) 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-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

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 Granulin in wild-type HAP1 cells as signal was lost in GRN (Granulin) 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

All lanes:

Western blot - Anti-Granulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) 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

• IP

Supplier Data

Immunoprecipitation - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (AB225532)

Granulin 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

All lanes:

Immunoprecipitation - Anti-Granulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>)

Predicted band size: 64 kDa

false