Anti-NSMase2 antibody [EPR29180-87]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSMase2 antibody [EPR29180-87] (AB322660)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NSMase2 with ab322660 at 1/100 (4.62 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human cerebrum.
The section was incubated with ab322660 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-NSMase2 antibody [EPR29180-87] (AB322660)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling NSMase2 with ab322660 at 1/100 (4.62 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human skeletal muscle (PMID : 10823942).
The section was incubated with ab322660 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-NSMase2 antibody [EPR29180-87] (AB322660)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling NSMase2 with ab322660 at 1/2000 (0.231 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse kidney (PMID : 10823942).
The section was incubated with ab322660 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-NSMase2 antibody [EPR29180-87] (AB322660)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NSMase2 with ab322660 at 1/2000 (0.231 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse cerebrum.
The section was incubated with ab322660 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

• WB

Supplier Data

Western blot - Anti-NSMase2 antibody [EPR29180-87] (AB322660)

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

Negative control : HEK-293(PMID : 10823942), B16-F0.

This antibody was tested on frozen and fresh Neuro-2a whole cell lysate and it was concluded that NSMase2 is the easy target for degradation.

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

To minimize protein degradation, cells (lanes 5) 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.

Exposure time : Lanes 1-4 : 180 seconds; Lane 5 : 114 seconds

All lanes:

Western blot - Anti-NSMase2 antibody [EPR29180-87] (ab322660) at 1/1000 dilution

Lane 1:

IMR-32 ( human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lanes 3 and 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

B16-F0 (mouse melanoma epithelial-like 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 1/100000 dilution

Observed band size: 71 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-NSMase2 antibody [EPR29180-87] (AB322660)

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

This antibody was tested on fresh and frozen IMR-32 whole cell lysate and it was concluded that NSMase2 is the easy target for degradation.

To minimize protein degradation, cells (lanes 1) 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-NSMase2 antibody [EPR29180-87] (ab322660) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) whole cell fresh lysate at 20 µg

Lane 2:

IMR-32 (human neuroblastoma neuroblast) 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: 71 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-NSMase2 antibody [EPR29180-87] (AB322660)

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

Negative control : skeletal muscle(PMID : 10823942)

The bands beneath the target band and above 30KD are likely to be degraded target fragments.

The identity of the bands below 20kDa are unknown.

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

All lanes:

Western blot - Anti-NSMase2 antibody [EPR29180-87] (ab322660) at 1/1000 dilution

Lane 1:

Human Midbrain tissue lysate at 20 µg

Lane 2:

Human striatum tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 71 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NSMase2 antibody [EPR29180-87] (AB322660)

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

Negative control : skeletal muscle, spleen, kidney(PMID : 10823942)

Low expression : cerebellum(PMID : 29867196)

The expression profile observed is consistent with what has been described in the literature (PMID : 29867196).

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

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

Exposure time : Lanes 1-4 : 180 seconds; Lanes 5-7 : 70 seconds

"

All lanes:

Western blot - Anti-NSMase2 antibody [EPR29180-87] (ab322660) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Mouse striatum tissue lysate at 20 µg

Lane 6:

Mouse midbrain tissue lysate at 20 µg

Lane 7:

Mouse 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 1/100000 dilution

Observed band size: 71 kDa,36 kDa

false