Anti-Lipoprotein lipase antibody [EPR31098-680]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized
293T (human embryonic kidney epithelial cell) cells transfected with a human LPL expression vector containing a myc-His-tag® (middle)
293T cells transfected with an empty expression vector containing a myc-His-tag® (Right)
labelling with ab328031 at 1/5000 dilution (0.01ug) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) transfected with an empty expression vector containing a myc-His-tag® and 293T cells transfected with a human LPL protein expression vector containing a myc-His-tag® labelling with ab328031 at 1/500 (1.044 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 positive staining in 293T cells transfected with a human LPL protein expression vectors containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain Myc at 1/100 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunohistochemical analysis of paraffin-embedded Mouse white adipose tissue labeling with ab328031 at 1/2000 (0.261 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining in mouse white adipose (PMID : 24899625).

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-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunohistochemical analysis of paraffin-embedded
(A) HEK-293T cells transfected with a mouse LPL expression vector containing a myc-His tag cell pellet
(B) HEK-293T cells transfected with a mouse LIPG expression vector containing a myc-His tag cell pellet
(C) HEK-293T cells transfected with a mouse LIPC expression vector containing a myc-His tag cell pellet
(D) HEK-293T cells transfected with an empty vector containing a myc-His tag cell pellet
labeling with ab328031 at 1/2000 (0.261 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining HEK-293T cells transfected with a mouse LPL expression vector containing a myc-His tag cell pellet (A); No staining in HEK-293T cells transfected with a mouse LIPG expression vector containing a myc-His tag cell pellet (B); No staining in HEK-293T cells transfected with a mouse LIPC expression vector containing a myc-His tag cell pellet (C); No staining in HEK-293T cells transfected with an empty plasmid vector containing a myc-His tag cell pellet (D).

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-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunohistochemical analysis of paraffin-embedded (A) L-929 (mouse connective tissue fibroblast) cell pellet and (B) Neuro-2a (mouse neuroblastoma neuroblast) cell pellet tissue labeling Lipoprotein lipase with ab328031 at 1/2000 (0.261 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining in L-929 cell pellet (A); Nearly no staining in Neuro-2a cell pellet (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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling with ab328031 at 1/2000 (0.261 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining in mouse cardiac muscle (PMID : 30598475).

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-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling with ab328031 at 1/2000 (0.261 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : Almost no staining in mouse spleen (PMID : 30598475).

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.

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized L-929 (mouse connective tissue fibroblast, Right) and Neuro-2a (mouse neuroblastoma neuroblast, Left) labelling with ab328031 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : Neuro-2a.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized
293T (human embryonic kidney epithelial cell) cells transfected with a mouse LPL expression vector containing a myc-His-tag® (Upper middle)
293T cells transfected with a mouse LIPG expression vector containing a myc-His-tag® (Upper right)
293T cells transfected a mouse LIPC expression vector containing a myc-His-tag® (Lower left)
293T cells transfected with an empty expression vector containing a myc-His-tag® (Lower middle)
labelling with ab328031 at 1/5000 dilution (0.01ug) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized L-929 (mouse connective tissue fibroblast) and Neuro-2a (mouse neuroblastoma neuroblast) cells labelling with ab328031 at 1/500 (1.044 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 cytoplasmic staining in L-929 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : Neuro-2a.
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).

• WB

Lab

Western blot - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (53KDa); Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Lipoprotein lipase antibody [EPR31098-680] (ab328031) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T cells transfected with a human Lipoprotein lipase 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='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 53 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Lipoprotein lipase antibody [EPR31098-680] (AB328031)

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

Low expression : Neuro-2a.

The identity of the bands lower than 37 kDa are unknown.

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

Exposure time : Lanes 1-2 : 180 seconds; Lanes 3-6 : 26 seconds.

All lanes:

Western blot - Anti-Lipoprotein lipase antibody [EPR31098-680] (ab328031) at 1/1000 dilution

Lane 1:

Mouse heart tissue lysate at 20 µg

Lane 2:

Mouse white fat tissue lysate at 20 µg

Lane 3:

L-929 (mouse connective tissue fibroblast) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

Undifferentiated 3T3-L1 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 6:

3T3-L1 differentiated for 6 days whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 53 kDa,36 kDa

false