Anti-Progerin antibody [EPR28694-72] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progerin antibody [EPR28694-72] - BSA and Azide free (AB320643)

This data was developed using ab320642, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Progerin with ab320642 at 1/500 (1.018 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human liver (PMID : 36497176).
The section was incubated with ab320642 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-Progerin antibody [EPR28694-72] - BSA and Azide free (AB320643)

This data was developed using ab320642, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human progerin expression vector containing a Flag-tag and (B) HEK-293T transfected with an empty vector containing a Flag-tag. tissue labeling Progerin with ab320642 at 1/500 (1.018 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) HEK-293T transfected with a human progerin expression vector containing a Flag-tag. No staining in (B) HEK-293T transfected with an empty vector containing a Flag-tag.
The section was incubated with ab320642 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-Progerin antibody [EPR28694-72] - BSA and Azide free (AB320643)

This data was developed using ab320642, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Progerin with ab320642 at 1/200 (2.545 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green).

Confocal image showing nuclear envelope staining in 293T cells transfected with human Progerin expression vector containing a Flag tag (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).

Anti-FLAG mouse monoclonal antibody was used to counterstain tubulin at 1/500 2ug/ml dilution (Magenta), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed antibody at 1/1000 2 ug/mL dilution. 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 2 ug/mL dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Progerin antibody [EPR28694-72] - BSA and Azide free (AB320643)

This data was developed using ab320642, the same antibody clone in a different buffer formulation.

Progerin was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) cells transfected with a human progerin expression vector containing a Flag-tag whole cell lysate with ab320642 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320642 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293T (human embryonic kidney epithelial cell) cells transfected with a human progerin expression vector containing a Flag-tag, whole cell lysate
Lane 2 : ab320642 IP in HEK-293T (human embryonic kidney epithelial cell) cells transfected with a human progerin expression vector containing a Flag-tag whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab320642 in HEK-293T cells transfected with a human progerin expression vector containing a Flag-tag whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Progerin antibody [EPR28694-72] (<a href='/en-us/products/primary-antibodies/progerin-antibody-epr28694-72-ab320642'>ab320642</a>) at 1/30 dilution

All lanes:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with a human progerin expression vector containing a Flag-tag whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-Progerin antibody [EPR28694-72] - BSA and Azide free (AB320643)

This data was developed using ab320642, the same antibody clone in a different buffer formulation.

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

Western blot analysis of HEK-293T cells transfected with a human progerin expression vector containing a Flag-tag whole cell lysate 2 ug mixed with HeLa whole cell lysate 20 ug. The membrane was cut into strips and each strip was incubated separately with the following antibodies : Lane 1 : anti Lamin A + Lamin C antibody ab108595 (1 : 1000), Lane 2 : anti Flag-tag antibody (1 : 10000), Lane 3 : anti-progerin ab320642 (1 : 1000).

The anti-progerin antibody ab320642 specifically detects human progerin.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 12714972, PMID : 33408413, PMID : 34694158).

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

In Western blot, Anti-Lamin A + Lamin C antibody - (ab108595) staining at 64-76KDa dilution.

In Western blot, Anti-Flag tag staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Progerin antibody [EPR28694-72] (<a href='/en-us/products/primary-antibodies/progerin-antibody-epr28694-72-ab320642'>ab320642</a>) at 1/1000 dilution

All lanes:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with a human progerin expression vector containing a Flag-tag whole cell lysate 2 μg mixed with HeLa (human cervical adenocarcinoma 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 1/100000 dilution

Observed band size: 74 kDa,64-76 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Progerin antibody [EPR28694-72] - BSA and Azide free (AB320643)

This data was developed using ab320642, 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 human Lamin A and Lamin C.

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

In Western blot, Anti-Lamin A + Lamin C antibody - (ab108595) staining at 64-76KDa dilution.

In Western blot, Anti-Flag tag staining at 1/10000 dilution.

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

All lanes:

Western blot - Anti-Progerin antibody [EPR28694-72] (<a href='/en-us/products/primary-antibodies/progerin-antibody-epr28694-72-ab320642'>ab320642</a>) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a DDDDK-tag, whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a human progerin expression vector containing a Flag-tag, whole cell lysate at 20 µg

Lane 3:

HEK-293T cells transfected with a human Lamin C expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 4:

HEK-293T cells transfected with a human Lamin A expression vector containing a myc-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: 74 kDa,36 kDa,64-76 kDa

false

Exposure time: 10s