Anti-UBQLN2 antibody [6H9]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [6H9] (AB190283)

Immunofluorescent analysis of HeLa cells labeling UBQLN2 with ab190283 at 1/1000 dilution. Vimentin is stained in red.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [6H9] (AB190283)

NIH/3T3 (mouse embyro fibroblast cell line) cells stained for UBQLN2 (green) using ab190283 at 1/1000 dilution in ICC/IF. Costained with chicken pAb to lamin A/C at 1/5,000 dilution (red). The blue is DAPI staining of nuclear DNA.

• WB

Supplier Data

Western blot - Anti-UBQLN2 antibody [6H9] (AB190283)

All lanes:

Western blot - Anti-UBQLN2 antibody [6H9] (ab190283) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embyro fibroblast cell line) cell lysate

Lane 2:

C6 (rat glial tumor cell line) cell lysate

Lane 3:

HEK-293 (human epithelial cell line from embryonic kidney) cell lysate

Lane 4:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

Lane 5:

SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate

Lane 6:

Rat whole brain lysate

Lane 7:

Mouse whole brain lysate

Predicted band size: 66 kDa

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• WB

Supplier Data

Western blot - Anti-UBQLN2 antibody [6H9] (AB190283)

All lanes:

Western blot - Anti-UBQLN2 antibody [6H9] (ab190283) at 1/1000 dilution

Lane 1:

Primary mouse neuron and glia cell cultures

Lane 2:

Primary mouse neuron and glia cells transduced with Human ubiquilin 2 wild type

Lane 3:

Primary mouse neuron and glia cells transduced with ubiquilin 2 P506T mutant

Lane 4:

Primary mouse neuron and glia cells transduced with ubiquilin 2 P497S mutant

Lane 5:

Primary mouse neuron and glia cell cultures transduced with enhanced GFP control

Lane 6:

HeLa cell lysate

Lane 7:

3T3 cell lysate

Predicted band size: 66 kDa

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• WB

Lab

Western blot - Anti-UBQLN2 antibody [6H9] (AB190283)

Western blot : Anti-UBQLN2 antibody [6H9] (ab190283) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab190283 was shown to bind specifically to UBQLN2. A band was observed at 73 kDa in wild-type A549 cell lysates with no signal observed at this size in UBQLN2 knockout cell line. To generate this image, wild-type and UBQLN2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-UBQLN2 antibody [6H9] (ab190283) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

UBQLN2 knockout A549 cell lysate at 20 µg

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

HEK293 Nuclear cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 73 kDa

false

• WB

Lab

Western blot - Anti-UBQLN2 antibody [6H9] (AB190283)

ab190283 was shown to react with UBQLN2 in wild-type HAP1 cells in Western blot with loss of signal observed in a UBQLN2 knockout cell line. Wild-type HAP1 and UBQLN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab190283 overnight at 4 °C at a 1/2000 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-UBQLN2 antibody [6H9] (ab190283) at 1/2000 dilution

Lane 1:

Wild-type HAP1 lysate at 20 µg

Lane 2:

UBQLN2 knock-out HAP1 lysate at 20 µg

Observed band size: 66 kDa

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