Anti-IDUA antibody antibody [EPR26926-251]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling IDUA with ab323701 at 1/100 (5.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney. The section was incubated with ab323701 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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-IDUA antibody antibody [EPR26926-251] (AB323701)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling IDUA with ab323701 at 1/100 (5.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on human skeletal muscle. The section was incubated with ab323701 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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-IDUA antibody antibody [EPR26926-251] (AB323701)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling IDUA with ab323701 at 1/100 (5.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human pancreas. The section was incubated with ab323701 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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-IDUA antibody antibody [EPR26926-251] (AB323701)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized IMR-90 (human lung fibroblast) cells labelling IDUA with ab323701 at 1/50 (10.2 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing lysosome staining in IMR-90 cells(shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : JAR.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab25630 Anti-LAMP1 mouse monoclonal antibody was used to counterstain tubulin at 1/200 2.5ug/ml 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 2ug/ml dilution.

Counter stain secondary antibody only control : Secondary antibody is ab150115 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 2ug/ml dilution.

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

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

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

HeLa transfected with siRNA specifically targeting IDUA whole cell lysate at 50 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: 60-90 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

The identity of the bands between 15 kDa and 50 kDa are unknown.

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

Human pancreas tissue lysate at 50 µg

Lane 2:

Human breast tissue lysate at 50 µg

Lane 3:

Human colon tissue lysate at 50 µg

Lane 4:

Human hippocampus tissue lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 60-90 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

Low expression : JEG-3, JAR

The identity of the bands between 15 kDa and 37 kDa are unknown.

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

Exposure time : Lane 1 : 3 seconds, Lanes 2-5 : 10 seconds, Lane 6 : 70 seconds

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

IMR-90 (human lung fibroblast) whole cell lysate at 50 µg

Lane 2:

U266B1 (human multiple myeloma B lymphocyte) whole cell lysate at 50 µg

Lane 3:

JEG-3 (human placenta epithelial cell) whole cell lysate at 50 µg

Lane 4:

JAR (human placenta choriocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 5:

HepG2 (human hepatocelullar carcinoma epithelial cell) whole cell lysate at 50 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: 60-90 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

Low expression : F9, MEF

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

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) whole cell lysate at 50 µg

Lane 2:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

MEF (mouse embryo fibroblast) whole cell lysate at 50 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: 60-90 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

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

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

mIMCD3 transfected with siRNA specifically targeting IDUA 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: 60-90 kDa,36 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

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

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

Rat testis tissue lysate at 20 µg

Lane 2:

Rat spleen tissue 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: 60-90 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

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.

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

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

McA-RH7777 (rat liver epithelial cell) whole cell lysate at 20 µg

Lane 2:

NMuLi (mouse liver epithelial cell) 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: 60-90 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

Low expression : skeletal muscle

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

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

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

Rat lung tissue lysate at 20 µg

Lane 2:

Rat skeletal muscle tissue lysate at 20 µg

Lane 3:

Rat kidney tissue 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: 60-90 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (AB323701)

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

The identity of the bands between 15 kDa and 50 kDa are unknown.

Exposure time : Lanes 1, 3-4 : 81 seconds, Lane 2 : 103 seconds

All lanes:

Western blot - Anti-IDUA antibody antibody [EPR26926-251] (ab323701) at 1/1000 dilution

Lane 1:

Mouse pancreas tissue lysate at 50 µg

Lane 2:

Rat pancreas tissue lysate at 50 µg

Lane 3:

Rat colon tissue lysate at 50 µg

Lane 4:

Rat brain tissue lysate at 50 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: 60-90 kDa

false