Anti-FGF21 antibody [EPR8314(2)]

• WB

Unknown

Western blot - Anti-FGF21 antibody [EPR8314(2)] (AB171941)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/5000 dilution

All lanes:

Human fetal liver tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

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• WB

Lab

Western blot - Anti-FGF21 antibody [EPR8314(2)] (AB171941)

False colour image of Western blot : Anti-FGF21 antibody [EPR8314(2)] staining at 1/1000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab171941 was shown to bind specifically to FGF21. A band was observed at 21 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in FGF21 knockout cell line ab265974 (knockout cell lysate ab256915). To generate this image, wild-type and FGF21 knockout HeLa 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 before development and with high-sensitivity chemiluminescence substrate and imaged with 16 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed at 1/20000 dilution.

All lanes:

Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/1000 dilution

Lane 1:

Wild-type HeLa Treated BFA (5 ug/mL, 6 h) cell lysate at 20 µg

Lane 2:

FGF21 knockout HeLa Treated BFA (5 ug/mL, 6 h) cell lysate at 20 µg

Lane 3:

Wild-type HeLa Vehicle Control BFA (0 ug/mL, 6 h) cell lysate at 20 µg

Lane 4:

FGF21 knockout HeLa Vehicle Control BFA (0 ug/mL, 6 h) cell lysate at 20 µg

Lane 5:

HepG2 Treated BFA (5 ug/mL, 6 h) cell lysate at 20 µg

Lane 6:

HepG2 cell lysate at 20 µg

Lane 7:

Empty

Lane 8:

Human Liver cell lysate at 10 µg

Secondary

All lanes:

HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) at 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 21 kDa

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• WB

Unknown

Western blot - Anti-FGF21 antibody [EPR8314(2)] (AB171941)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-FGF21 antibody [EPR8314(2)] (ab171941) at 1/5000 dilution

All lanes:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

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