Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human adipose tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human adipose tissue.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining Hormone sensitive lipase with ab325336 at a 1/2000 (0.248 μg/ml) dilution, ab315800 anti-CYP51A1 used at a 1/2000 (0.26 μg/ml) dilution, and ab270534 anti-DDX4 used at a 1/2000 (0.238 μg/ml) dilution.

Panel A : anti-Hormone sensitive lipase (green; Opal™520), anti-CYP51A1 (magenta; Opal™690), anti-DDX4 (gray; Opal™570) on human testis.

Panel B : anti-Hormone sensitive lipase staining spermatid in human testis.

Panel C : anti-CYP51A1 staining leydig cells in human testis.

Panel D : anti-DDX4 staining spermatogonia in human testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325336, ab315800 and ab270534 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human breast tissue staining Hormone sensitive lipase with ab325336 at a 1/2000 (0.248 μg/ml) dilution, ab315240 anti-Perlecan used at a 1/2000 (0.254 μg/ml) dilution, and ab180862 anti-SOX10 used at a 1/4000 (0.03 μg/ml) dilution.

Panel A : anti-Hormone sensitive lipase (green; Opal™520), anti-Perlecan (magenta; Opal™690), anti-SOX10 (gray; Opal™570) on human breast.

Panel B : anti-Hormone sensitive lipase staining adipocytes in human breast.

Panel C : anti-Perlecan staining basement membrane and endothelium in human breast.

Panel D : anti-SOX10 staining nucleus in human breast.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325336, ab315240 and ab180862 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte);Huh7 (human hepatocellular carcinoma epithelial cell) cells labelling Hormone sensitive lipase/HSL with ab325336 at 1/1000 (0.496 μg/ml) dilution , followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 dilution (Green).

Confocal image showing cytoplasmic staining in Raji cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Negative control : Huh7.

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).

Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human breast tissue staining Hormone sensitive lipase with ab325336 at a 1/2000 (0.248 μg/ml) dilution, ab318972 anti-COL8A1 used at a 1/2000 (0.264 μg/ml) dilution, and ab180862 anti-SOX10 used at a 1/4000 (0.03 μg/ml) dilution.

Panel A : anti-Hormone sensitive lipase (green; Opal™520), anti-COL8A1 (magenta; Opal™690), anti-SOX10 (gray; Opal™570) on human breast.

Panel B : anti-Hormone sensitive lipase staining adipocytes in human breast.

Panel C : anti-COL8A1 staining basement membrane in human breast.

Panel D : anti-SOX10 staining nucleus in human breast.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325336, ab318972 and ab180862 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized undifferentiated 3T3-L1 (mouse embryonic fibroblast);differentiated 3T3-L1 cells labelling Hormone sensitive lipase/HSL with ab325336 at 1/1000 (0.496 μg/ml) dilution , followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 dilution (Green).

Confocal image showing no staining in undifferentiated 3T3-L1 cells, showing cytoplasmic staining in differentiated 3T3-L1 cells(shown in magenta). The counterstain was observed in green. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Lipid Droplets Green Fluorescence Assay Kit with BODIPY493/503 was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on spermatid of mouse testis.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse breast tissue staining Hormone sensitive lipase/HSL with ab325336 at a 1/2000 (0.248 μg/ml) dilution, ab324191 anti-E Cadherin used at a 1/500 (2.088 μg/ml) dilution and ab180862 anti-SOX10 used at a 1/4000 (0.03 μg/ml) dilution.

Panel A : anti-Hormone sensitive lipase/HSL (green; Opal™520), anti-E Cadherin (magenta; Opal™690), anti-SOX10 (gray; Opal™570) on mouse breast.

Panel B : anti-Hormone sensitive lipase/HSL staining adipocytes in mouse breast.

Panel C : anti-E Cadherin staining epithelium in mouse breast.

Panel D : anti-SOX10 staining on nucleus in mouse breast.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325336, ab324191 and ab180862 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse adipose tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse adipose tissue.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Lab

Western blot - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

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

Negative control : lung, liver, spleen.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 8496671).

The identity of the bands higher than 100 kDa and lower than 75 kDa are unknown.

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

All lanes:

Western blot - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] (<a href='/products/primary-antibodies/hormone-sensitive-lipase-hsl-antibody-epr27268-59-ab325336'>ab325336</a>) at 1/1000 dilution

Lane 1:

Human adrenal gland tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human spleen 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: 84 kDa,36 kDa

false

Exposure time: 59s

• WB

Lab

Western blot - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

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

Negative control : Huh7.

The identity of the bands higher than 100 kDa and lower than 75 kDa are unknown.

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

All lanes:

Western blot - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] (<a href='/products/primary-antibodies/hormone-sensitive-lipase-hsl-antibody-epr27268-59-ab325336'>ab325336</a>) at 1/1000 dilution

Lane 1:

Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 2:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 83 kDa,36 kDa

false

Exposure time: 59s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human liver (PMID : 8496671).

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse liver.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human lung (PMID : 8496671).

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hormone sensitive lipase/HSL antibody [EPR27268-59] - BSA and Azide free (AB325348)

This data was developed using ab325336, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Hormone sensitive lipase/HSL with ab325336 at 1/2000 (0.248 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse lung.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins