All lanes : Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : SLC27A4 knockout HEK293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HEK-293 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 72 kDa
Observed band size: 72 kDa

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.

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 Fluor^® 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.

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

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 3 : HEK293 (Human embryonic kidney) whole cell lysate
Lane 4 : Human testis lysate

Lysates/proteins at 20 µg per lane.

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


Exposure time: 1 minute

Blocking/dilution buffer: 5% NFDM/TBST.

All lanes : Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

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


Exposure time: 30 seconds

Blocking/dilution buffer: 5% NFDM/TBST.

All lanes : Anti-SLC27A4 / FATP4 antibody [EPR17319-26] (ab200353) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat kidney lysate
Lane 7 : Rat spleen lysate
Lane 8 : C6 (Rat glial tumor cells) whole cell lysate
Lane 9 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 10 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 11 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

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


Exposure time: 5 seconds

Blocking/dilution buffer: 5% NFDM/TBST.