Anti-SLC27A4 / FATP4 antibody [EPR17319] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC27A4 / FATP4 antibody [EPR17319] - BSA and Azide free (AB271965)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SLC27A4 / FATP4 with ab199719 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human kidney 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).

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab199719).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SLC27A4 / FATP4 antibody [EPR17319] - BSA and Azide free (AB271965)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling SLC27A4 / FATP4 with ab199719 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm staining on HepG2 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor^®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows : -
-ve control 1 : ab199719 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) 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 (ab199719).

• IP

Supplier Data

Immunoprecipitation - Anti-SLC27A4 / FATP4 antibody [EPR17319] - BSA and Azide free (AB271965)

SLC27A4 / FATP4 was immunoprecipitated from 1mg of HEK293 (Human embryonic kidney) whole cell lysate with ab199719 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab199719 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HEK293 whole cell lysate 10μg (Input). Lane 2 : HEK293 whole cell lysate following IP. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199719 in HEK293 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure is 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab199719).

All lanes:

Immunoprecipitation - Anti-SLC27A4 / FATP4 antibody [EPR17319] - C-terminal (<a href='/en-us/products/primary-antibodies/slc27a4-fatp4-antibody-epr17319-c-terminal-ab199719'>ab199719</a>)

Predicted band size: 72 kDa

Observed band size: 72 kDa

false

• WB

Supplier Data

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319] - BSA and Azide free (AB271965)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : SLC27A4/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 - ab199719 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab199719 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. ab199719 and ab8245 (loading control to GAPDH) were diluted 1/2000 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h 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 (ab199719).

All lanes:

Western blot - Anti-SLC27A4 / FATP4 antibody [EPR17319] - C-terminal (<a href='/en-us/products/primary-antibodies/slc27a4-fatp4-antibody-epr17319-c-terminal-ab199719'>ab199719</a>)

Predicted band size: 72 kDa

false