Anti-SREBP1 antibody [EPR30539-645]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Mouse fatty liver tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse fatty liver. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized McA-RH7777 (rat liver epithelial cell) cells labelling SREBP1 with ab326106 at 1/50 (10.38 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear and cytoplasmic staining in McA-RH7777 cells(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).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326106 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse liver (PMID : 9062340). 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Mouse adrenal gland tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse adrenal gland (PMID : 9062340). 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse breast carcinoma. 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on rat liver (PMID : 16554040). 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Rat adrenal gland tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on rat adrenal gland (PMID : 16554040). 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression tissue : weak staining on scattered cells of 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling SREBP1 with ab326106 at 1/200 (2.595 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression tissue : almost no 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunohistochemical analysis of paraffin-embedded (A) Undifferentiated 3T3-L1 (mouse embryonic fibroblast) cell pellet (B) 3T3-L1 cell pellet differentiated for 6 days tissue labeling SREBP1 with ab326106 at 1/100 (5.19 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Weak staining on scattered cells of undifferentiated 3T3-L1 cell pellet (image A), with increased expression of SREBF1 in 3T3-L1 cell pellet differentiated for 6 days (image B). 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

• WB

Lab

Western blot - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

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

Samples are non-boiled as boiling may cause protein aggregation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 24776759).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1 : 70 seconds, Lanes 2-4 : 15 seconds

All lanes:

Western blot - Anti-SREBP1 antibody [EPR30539-645] (ab326106) at 1/1000 dilution

Lane 1:

Rat adrenal gland tissue lysate at 50 µg

Lane 2:

Rat liver tissue lysate at 50 µg

Lane 3:

Rat spleen tissue lysate at 50 µg

Lane 4:

McA-RH7777 (rat liver 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: 65(matureform) kDa,113-120(alternativesplicing) kDa,36 kDa

false

• WB

Lab

Western blot - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

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

Samples are non-boiled as boiling may cause protein aggregation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 24776759).

This antibody could detect both SREBP-1c and SREBP-1a.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-2 : 70 seconds, Lanes 3-4 : 15 seconds

All lanes:

Western blot - Anti-SREBP1 antibody [EPR30539-645] (ab326106) at 1/1000 dilution

Lane 1:

Untreated 3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 50 µg

Lane 2:

3T3-L1 differentiation for 6days whole cell lysate at 50 µg

Lane 3:

Mouse liver tissue lysate at 50 µg

Lane 4:

Mouse spleen 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: 65(matureform) kDa,113-120(alternativesplicing) kDa,36 kDa

false

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SREBP1 antibody [EPR30539-645] (AB326106)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat splenocyte cells labelling SREBP1 with ab326106 at 1/50 (10.38 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Negative control : Confocal image showing no staining in rat splenocytes (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).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326106 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.