Anti-GOLPH4/GPP130 antibody [EPR13439]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH4/GPP130 antibody [EPR13439] (AB197595)

Immunohistochemical analysis of paraffin-embedded Human renal adenocarcinoma tissue labeling GOLPH4/GPP130 with ab197595 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on Human renal adenocarcinoma tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• WB

Supplier Data

Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (AB197595)

Blocking/Dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (PMID : )20130081 and 9201717

All lanes:

Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/5000 dilution

Lane 1:

HepG2(Human liver hepatocellular carcinoma) whole cell lysate at 10 µg

Lane 2:

293(Human 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: 82 kDa

Observed band size: 130 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (AB197595)

Blocking/Dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID : 20130081 and PMID : 9201717

All lanes:

Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/2500 dilution

All lanes:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 82 kDa

Observed band size: 130 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (AB197595)

Lanes 1-4 : Merged signal (red and green). Green - ab197595 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.

ab197595 Anti-GOLPH4/GPP130 antibody [EPR13439] was shown to specifically react with GOLPH4/GPP130 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266123 (knockout cell lysate ab257455) was used. Wild-type and GOLPH4/GPP130 knockout samples were subjected to SDS-PAGE. ab197595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

GOLIM4 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human GOLIM4 (GOLPH4/GPP130) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-golim4-golph4-gpp130-knockout-hek-293t-cell-line-ab266123'>ab266123</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MOLT-4 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: 82 kDa

Observed band size: 130 kDa

false