Anti-GOLPH2 antibody [OTI6C9]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Paraffin-embedded human prostate tissue stained for GOLPH2 using ab119800 at 1/150 dilution in immunohistochemical analysis.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Paraffin-embedded human endometrium adenocarcinoma tissue stained for GOLPH2 using ab119800 at 1/150 dilution in immunohistochemical analysis.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Paraffin-embedded human thyroid carcinoma tissue stained for GOLPH2 using ab119800 at 1/150 dilution in immunohistochemical analysis.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Paraffin-embedded human endometrium tissue stained for GOLPH2 using ab119800 at 1/150 dilution in immunohistochemical analysis.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Paraffin-embedded human bladder carcinoma tissue stained for GOLPH2 using ab119800 at 1/150 dilution in immunohistochemical analysis.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Overlay histogram showing HeLa cells stained with ab119800 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab119800, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

Paraffin-embedded human ovary adenocarcinoma tissue stained for GOLPH2 using ab119800 at 1/150 dilution in immunohistochemical analysis.

• WB

Lab

Western blot - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

False colour image of Western blot : Anti-GOLPH2 antibody [OTI6C9] staining at 1/500 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab119800 was shown to bind specifically to GOLPH2. A band was observed at 70-75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in GOLM1 knockout cell line ab266505 (knockout cell lysate ab257976). To generate this image, wild-type and GOLM1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-GOLPH2 antibody [OTI6C9] (ab119800) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 10 µg

Lane 2:

GOLM1 knockout HEK-293T cell lysate at 10 µg

Lane 2:

Western blot - Human GOLM1 (GOLPH2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-golm1-golph2-knockout-hek-293t-cell-line-ab266505'>ab266505</a>)

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Lane 5:

MDA-MB-231 cell lysate at 20 µg

Lane 6:

LNCaP cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 70-75 kDa

false

• WB

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Western blot - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

HEK293T cell lysates were generated from transient transfection of the cDNA clone (RC200086)

All lanes:

Western blot - Anti-GOLPH2 antibody [OTI6C9] (ab119800) at 1/500 dilution

Lane 1:

HepG2 (human liver hepatocellular carcinoma cell line) cell extract at 35 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell extract at 35 µg

Lane 3:

SVT2 cell extract at 35 µg

Lane 4:

A549 (human lung carcinoma cell line) cell extract at 35 µg

Lane 5:

COS-7 (african green monkey kidney fibroblast-like cell line) cell extract at 35 µg

Lane 6:

Jurkat (human T cell leukemia cell line from peripheral blood) cell extract at 35 µg

Lane 7:

MDCK (canine kidney cell line) cell extract at 35 µg

Lane 8:

PC-12 (rat adrenal gland pheochromocytoma cell line) cell extract at 35 µg

Lane 9:

MCF7 (human breast adenocarcinoma cell line) cell extract at 35 µg

Predicted band size: 45 kDa

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

COS-7 (african green monkey kidney fibroblast-like cell line) cells transiently transfected by pCMV6-ENTRY GOLPH2 stained for GOLPH2 (green) using ab119800 at 1/100 dilution in ICC/IF.

• WB

Unknown

Western blot - Anti-GOLPH2 antibody [OTI6C9] (AB119800)

All lanes:

Western blot - Anti-GOLPH2 antibody [OTI6C9] (ab119800) at 1/500 dilution

Lane 1:

HEK293T cell lysate transfected with pCMV6-ENTRY control at 5 µg

Lane 2:

HEK293T cell lysate transfected with pCMV6-ENTRY GOLPH2 at 5 µg

Predicted band size: 45 kDa

false