Anti-SRSF5 antibody

• WB

Unknown

Western blot - Anti-SRSF5 antibody (AB67175)

All lanes:

Western blot - Anti-SRSF5 antibody (ab67175) at 1/500 dilution

All lanes:

human liver tissue lysate at 25 µg/mL

Secondary

All lanes:

Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

Predicted band size: 31 kDa

Observed band size: 39 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SRSF5 antibody (AB67175)

ab67175, at 1.5μg/ml, staining SRSF5 in human ovary by immunohistochemistry using formalin-fixed, paraffin-embedded tissue.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SRSF5 antibody (AB67175)

ICC/IF image of ab67175 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab67175, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• WB

Unknown

Western blot - Anti-SRSF5 antibody (AB67175)

All lanes:

Western blot - Anti-SRSF5 antibody (ab67175) at 1/500 dilution

Lane 1:

SRSF5 transfected 293T cell lysate at 25 µg/mL

Lane 2:

Non-transfected 293T cell lysate at 25 µg/mL

Secondary

All lanes:

Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

Predicted band size: 31 kDa

Observed band size: 27 kDa,39 kDa

false

• WB

Lab

Western blot - Anti-SRSF5 antibody (AB67175)

Lanes 1 - 4 : Merged signal (red and green). Green - ab67175 observed at 40 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab67175 was shown to react with SRSF5 in wild-type HEK-293T cells in Western blot with loss of signal observed in SRSF5 knockout cell line ab266246 (SRSF5 knockout cell lysate 258704). Wild-type HEK-293T and SRSF5 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab67175 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SRSF5 antibody (ab67175) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SRSF5 knockout HEK-293T cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 31 kDa

Observed band size: 40 kDa

false

• WB

Lab

Western blot - Anti-SRSF5 antibody (AB67175)

Lanes 1 - 4 : Merged signal (red and green). Green - ab67175 observed at 40 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab67175 was shown to react with SRSF5 in wild-type HEK-293T cells in Western blot with loss of signal observed in SRSF5 knockout cell line ab266246 (SRSF5 knockout cell lysate ab258704). Wild-type HEK-293T and SRSF5 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab67175 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SRSF5 antibody (ab67175) at 1/500 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SRSF5 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SRSF5 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-srsf5-knockout-hek-293t-cell-line-ab266246'>ab266246</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 31 kDa

Observed band size: 40 kDa

false