Anti-HuR / ELAVL1 antibody [19F12AE12]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HuR / ELAVL1 antibody [19F12AE12] (AB170193)

Immunocytochemistry on HeLa cells.

Cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15minutes). The cells were then incubated with the anti-HuR antibody (5μg/mL) for 2 hours at room temperature or over night at 4°C. The secondary antibody used was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. A cytoplasmic antibody (ab75834) was used as counter stain (green). 10% goat serum was used as a blocking agent for all blocking steps.

The target protein localizes mainly to the nucleus.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HuR / ELAVL1 antibody [19F12AE12] (AB170193)

Immunocytochemistry on SH-SY5Y cells (Human neuroblastoma cell line).

Cells cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were then incubated with the anti-HuR antibody (5μg/mL) for 2 hours at room temperature or overnight at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. A cytoplasmic antibody (ab75834) was used as counter stain (Green). 10% Goat serum was used as a blocking agent for all blocking steps.

The target protein localizes mainly to the nucleus.

• Flow Cyt

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Flow Cytometry - Anti-HuR / ELAVL1 antibody [19F12AE12] (AB170193)

Flow cytometric analysis of methanol-fixed HeLa cells labeling HuR / ELAVL1 with ab170193 at 1μg/mL (blue) or an equal amount of an isotype control antibody (red).

• WB

Lab

Western blot - Anti-HuR / ELAVL1 antibody [19F12AE12] (AB170193)

Lanes 1 - 4 : Merged signal (red and green). Green - ab170193 observed at 36 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab170193 was shown to react with ELAVL1 in wild-type SW480 cells in western blot. Loss of signal was observed when ELAVL1 knockout sample was used. Wild-type and ELAVL1 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBS-T (0.1% Tween^®) before incubation with ab170193 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 2 μg/ml 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 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HuR / ELAVL1 antibody [19F12AE12] (ab170193) at 2 µg/mL

Lane 1:

Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 40 µg

Lane 2:

ELAVL1 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 40 µg

Lane 2:

Western blot - Human ELAVL1 knockout SW480 cell line (<a href='/en-us/products/cell-lines/human-elavl1-knockout-sw480-cell-line-ab270485'>ab270485</a>)

Lane 3:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 4:

HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 36 kDa

Observed band size: 36 kDa

false

• WB

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Western blot - Anti-HuR / ELAVL1 antibody [19F12AE12] (AB170193)

All lanes:

Western blot - Anti-HuR / ELAVL1 antibody [19F12AE12] (ab170193) at 1 µg/mL

Lane 1:

H9C2 Rat Cell Lysate at 20 µg

Lane 2:

H4IIE Rat Cell Lysate at 20 µg

Lane 3:

MEF Mouse Cell Lysate at 20 µg

Lane 4:

3T3 Mouse Cell Lysate at 20 µg

Lane 5:

SH-SY5Y Cell Lysate at 20 µg

Lane 6:

HepG2 Cell Lysate at 20 µg

Secondary

All lanes:

Goat anti-mouse AP at 1/5000 dilution

Predicted band size: 36 kDa

true

• WB

Supplier Data

Western blot - Anti-HuR / ELAVL1 antibody [19F12AE12] (AB170193)

All lanes:

Western blot - Anti-HuR / ELAVL1 antibody [19F12AE12] (ab170193) at 1 µg/mL

Lane 1:

Human Liver Homogenate at 20 µg

Lane 2:

Human Heart Homogenate at 20 µg

Lane 3:

HepG2 Cell Lysate at 20 µg

Lane 4:

HeLa Cell Lysate at 20 µg

Lane 5:

SH-SY5Y Cell Lysate at 20 µg

Lane 6:

Jurkat Cell Lysate at 20 µg

Secondary

All lanes:

Goat anti-mouse IR800 at 1/5000 dilution

Predicted band size: 36 kDa

true