Anti-SLC27A4 / FATP4 antibody [EPR17319-26]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (AB200353)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling SLC27A4 / FATP4 with purified ab200353 at 1/250 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• WB

Supplier Data

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (AB200353)

Blocking/dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/5000 dilution

Lane 1:

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

Lane 2:

HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

Lane 3:

HEK293 (Human embryonic kidney) whole cell lysate at 20 µg

Lane 4:

Human testis lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 72 kDa

Observed band size: 72 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (AB200353)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : SLC27A/FATP4 knockout HAP1 cell lysate (20 μg)
Lane 3 : HEK293 cell lysate (20 μg)
Lane 4 : HepG2 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab200353 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab200353 was shown to specifically react with SLC27A4/FATP4 when SLC27A4/FATP4 knockout samples were used. Wild-type and SLC27A4/FATP4 knockout samples were subjected to SDS-PAGE. ab200353 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) andGoat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353)

Predicted band size: 72 kDa

false

• WB

Lab

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (AB200353)

Lanes 1-4 : Merged signal (red and green). Green - ab200353 observed at 72 kDa. Red - loading control ab8245 observed at 36 kDa.

ab200353 Anti-SLC27A4 / FATP4 antibody [EPR17319-26] was shown to specifically react with SLC27A4 / FATP4 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266114 (knockout cell lysate ab257677) was used. Wild-type and SLC27A4 / FATP4 knockout samples were subjected to SDS-PAGE. ab200353 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-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

SLC27A4 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human SLC27A4 (FATP4) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-slc27a4-fatp4-knockout-hek-293t-cell-line-ab266114'>ab266114</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HEK-293 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: 72 kDa

Observed band size: 72 kDa

false

• WB

Supplier Data

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (AB200353)

Blocking/dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

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: 72 kDa

Observed band size: 72 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (AB200353)

Blocking/dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat kidney lysate at 10 µg

Lane 7:

Rat spleen lysate at 10 µg

Lane 8:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 9:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 10:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 11:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 72 kDa

Observed band size: 72 kDa

false

Exposure time: 5s