Anti-Fyn antibody [25/FYN]

• WB

Supplier Data

Western blot - Anti-Fyn antibody [25/FYN] (AB300634)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The identity of the lower MW band at approximately 10kDa (in lane 1/2/3) is unknown. Exposure times : Lanes 1-4 : 92 seconds Lanes 5-6 : 3 minutes

All lanes:

Western blot - Anti-Fyn antibody [25/FYN] (ab300634) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Lane 2:

Ramos (human Burkitts lymphoma B lymphocyte), whole cell lysate at 10 µg

Lane 3:

C2C12 (mouse myoblasts), whole cell lysate at 10 µg

Lane 4:

Human thymus tissue lysate at 10 µg

Lane 5:

C6 (rat glial tumor cell), whole cell lysate at 10 µg

Lane 6:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 59 kDa

false

• WB

Supplier Data

Western blot - Anti-Fyn antibody [25/FYN] (AB300634)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS. False colour image of Western blot : Anti-Fyn antibody [25/FYN] (ab300634) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab300634 was shown to bind specifically to Fyn. A band was observed at 59 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in the Fyn knockout cell line. To generate this image, wild-type and Fyn knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in equal volume of 0.1 % TBS 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-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution. Performed under reducing conditions.

All lanes:

Western blot - Anti-Fyn antibody [25/FYN] (ab300634) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

Fyn knockout HEK-293, whole cell lysate at 20 µg

Lane 3:

Ramos (human Burkitts lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

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

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Observed band size: 59 kDa

false