Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) cells labelling SUN1 with ab323868 at 1/100 (5.1 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 nuclear envelope staining in C6 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 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling SUN1 with ab323868 at 1/1000 (0.51 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear envelope staining on mouse spleen.
The primary antibody was incubated 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-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SUN1 with ab323868 at 1/1000 (0.51 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear envelope staining on mouse cerebrum.
The primary antibody was incubated 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-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SUN1 with ab323868 at 1/1000 (0.51 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear envelope staining on rat kidney.
The primary antibody was incubated 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-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling SUN1 with ab323868 at 1/1000 (0.51 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear envelope staining on rat spleen.
The primary antibody was incubated 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh frozen) tissue labeling SUN1 with ab323868 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on rat spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323868 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using [Standard ab323868], the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh frozen) tissue labeling SUN1 with ab323868 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on mouse spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab323868 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh frozen) tissue labeling SUN1 with ab323868 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-SUN1 (ab323868 green) anti-NeuN (ab190565 magenta) on mouse cerebrum.
Panel B : anti-SUN1 stained on mouse cerebrum.
Panel C : anti-NeuN stained in neurons of mouse cerebrum.
The section was incubated in two rounds of staining : in the order of ab323868 and ab190565 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Rat cerebrum (fresh frozen) tissue labeling SUN1 with ab323868 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-SUN1 (ab323868 green) anti-NeuN (ab190565 magenta) on rat cerebrum.
Panel B : anti-SUN1 stained on rat cerebrum.
Panel C : anti-NeuN stained in neurons of rat cerebrum.
The section was incubated in two rounds of staining : in the order of ab323868 and ab190565 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling SUN1 with ab323868 at 1/100 (5.1 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 nuclear envelope 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 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.

• WB

Supplier Data

Western blot - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

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

The band above 250kda is likely to be SUN1 oligomers (PMID : 29813079).

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

All lanes:

Western blot - Anti-SUN1 antibody [EPR29563-76] (<a href='/en-us/products/primary-antibodies/sun1-antibody-epr29563-76-ab323868'>ab323868</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Neuro-2a transfected with siRNA specifically targeting SUN1 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: 84-101 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

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

The bands beneath the target band are likely to be degradation products.

The band above 250kda is likely to be SUN1 oligomers (PMID : 29813079).

In lane 1-3 the lysate was stored at -80°C prior to Western blotting. The bands beneath the target band (84 kDa) are likely to be degradation products. In lane 4-6 to minimize protein degradation tissues were lysed immediately after harvest and used 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-SUN1 antibody [EPR29563-76] (<a href='/en-us/products/primary-antibodies/sun1-antibody-epr29563-76-ab323868'>ab323868</a>) at 1/1000 dilution

Lane 1:

Mouse brain frozen tissue lysate at 20 µg

Lane 2:

Mouse testis frozen tissue lysate at 20 µg

Lane 3:

Mouse spleen frozen tissue lysate at 20 µg

Lane 4:

Mouse brain fresh tissue lysate at 20 µg

Lane 5:

Mouse testis fresh tissue lysate at 20 µg

Lane 6:

Mouse spleen fresh 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: 84-101 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

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

Low expression : skin(PMID : 20711465)

The bands beneath the target band are likely to be degradation products.

The band above 250kda is likely to be SUN1 oligomers (PMID : 29813079).

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

Exposure time : Lane1-2 : 48 seconds lane3-5 : 59 seconds lane6 : 37 seconds

All lanes:

Western blot - Anti-SUN1 antibody [EPR29563-76] (<a href='/en-us/products/primary-antibodies/sun1-antibody-epr29563-76-ab323868'>ab323868</a>) at 1/1000 dilution

Lane 1:

Rat embryo tissue lysate at 20 µg

Lane 2:

Rat skin tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat testis tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

Lane 6:

C6 (rat glial tumor glial 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: 84-101 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-SUN1 antibody [EPR29563-76] - BSA and Azide free (AB325950)

This data was developed using ab323868, the same antibody clone in a different buffer formulation.

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

This antibody does not cross-react with mouse SUN2 or SUN3.

The expression profile/ molecular weight observed is consistent with what has been described in the literature(PMID : 20711465). This antibody can recognize the different isoforms of SUN1.

The band above 250kda is likely to be SUN1 oligomers (PMID : 29813079).

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

In Western blot Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) - Loading Control (ab181602) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-SUN1 antibody [EPR29563-76] (<a href='/en-us/products/primary-antibodies/sun1-antibody-epr29563-76-ab323868'>ab323868</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a mouse SUN1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse SUN2 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a mouse SUN3 expression vector containing a His-tag, 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: 101 kDa,36 kDa

false

Exposure time: 8s