Anti-UBQLN2 antibody [EPR28592-74]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling UBQLN2 with ab323454 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly cytoplasmic staining in SH-SY5Y cell line and no staining in HL-60 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : HL-60.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type A549 (Human lung carcinoma epithelial cell) cell pellet /(B) UBQLN2 knockout A549 cell pellet labeling UBQLN2 with ab323454 at 1/8000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on wild-type A549 cell pellet (A), no staining on UBQLN2 knockout A549 cell pellet (B). The section was incubated with ab323454 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded SH-SY5Y (human neuroblastoma epithelial cell) tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on SH-SY5Y. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded HAP1 (human chronic myelogenous leukemia near-haploid cell) tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HAP1. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human glioma. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human ovarian carcinoma. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on human skeletal muscle. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling UBQLN2 with ab323454 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HAP1 (human chronic myelogenous leukemia near-haploid cell) cells labelling UBQLN2 with ab323454 at 1/100 (5.05 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly cytoplasmic staining in HAP1 cell line and no staining in FaDu cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : FaDu.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized UBQLN2 KO A549 (UBQLN2 knockout human lung carcinoma epithelial cell)/Wild-type A549 cells labelling UBQLN2 with ab323454 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly cytoplasmic staining in parental A549 cell line and no staining in UBQLN2 knockout A549 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on rat skeletal muscle. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : weak staining on mouse skeletal muscle. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling UBQLN2 with ab323454 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly cytoplasmic staining in PC-12 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling UBQLN2 with ab323454 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly cytoplasmic staining in Neuro-2a cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling UBQLN2 with ab323454 at 1/8000 (0.063 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum. The section was incubated with ab323454 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the lower MW band at approximately 20 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (ab323454) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

HeLa transfected with siRNA specifically targeting UBQLN2 whole cell lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa,36 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32413959, PMID : 35247097).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW band at approximately 20 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 6 seconds; Lanes 3-4 : 3 seconds.

All lanes:

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (ab323454) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a GAPDH loading control at a 1/200000 dilution. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32413959, PMID : 35247097). In Western blot, ab323454 was shown to bind specifically to UBQLN2. Target of interest was observed at 70 kDa in treated wild-type A549 cell lysates (lane 1) with no signal observed at this size in UBQLN2 knockout cell line (lane 2).

All lanes:

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (ab323454) at 1/1000 dilution

Lane 1:

Wild-type A549(Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

UBQLN2 knockout A549 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : FaDu, HL-60.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32413959, PMID : 35247097).

The bands beneath the target band are likely to be degraded target fragments.

The identity of the lower MW band at approximately 20 kDa in lane 9 is unknown.

Lanes 1-7 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 8-9 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-7 : 26 seconds; Lanes 8-9 : 92 seconds

All lanes:

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (ab323454) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

RPMI-8226 (human plasmacytoma B Lymphocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

786-O (human kidney epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 5:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 6:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg with NFDM/TBST

Lane 7:

FaDu (human pharyngeal carcinoma epithelial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 8:

Human liver tissue lysate at 20 µg with NFDM/TBST

Lane 9:

Human brainstem tissue lysate at 20 µg with NFDM/TBST

Secondary

Lanes 1 - 7:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Lanes 8 - 9:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 70 kDa,36 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Dot blot analysis of UBQLN2 using ab323454 at 1 : 1000 (0.505 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Purified 293T cells transfected with a human UBQL2 expression vector containing a His-tag, whole cell lysate
Lane2 : Purified 293T cells transfected with a human UBQL4 expression vector containing a His-tag, whole cell lysate
Lane3 : Purified 293T cells transfected with a human UBQL3 expression vector containing a His-tag, whole cell lysate
Lane4 : Purified 293T cells transfected with a human UBQL1 expression vector containing a His-tag, whole cell lysate

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human UBQL1, UBQL3 and UBQL4.

In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-UBQLN2 antibody [EPR28592-74] (ab323454) at 1/1000 dilution

Lane 1:

Purified 293T cells transfected with a human UBQL2 expression vector containing a His-tag, whole cell lysate

Lane 2:

Purified 293T cells transfected with a human UBQL4 expression vector containing a His-tag, whole cell lysate

Lane 3:

Purified 293T cells transfected with a human UBQL3 expression vector containing a His-tag, whole cell lysate

Lane 4:

Purified 293T cells transfected with a human UBQL1 expression vector containing a His-tag, whole cell lysate

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (AB323454)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : skeletal muscle.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 32413959, PMID : 35247097).

In lanes 1-7, the lysates was stored at -80°C prior to Western Blotting. The bands beneath the target band are likely to be degraded target fragments.

To minimize protein degradation, cells (lanes 8-10) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW band at approximately 20 kDa in lanes 2, 4 and 6 is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-9 : 3 seconds; Lane 10 : 1 second.

All lanes:

Western blot - Anti-UBQLN2 antibody [EPR28592-74] (ab323454) at 1/1000 dilution

Lanes 1 and 10:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg with NFDM/TBST

Lanes 2 and 9:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

Mouse brainstem tissue lysate at 20 µg with NFDM/TBST

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg with NFDM/TBST

Lane 6:

Rat brainstem tissue lysate at 20 µg with NFDM/TBST

Lane 7:

Rat skeletal muscle tissue lysate at 20 µg with NFDM/TBST

Lane 8:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa,36 kDa

false