Anti-Adipose Triglyceride Lipase antibody [EPR19650]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

IHC image of Adipose Triglyceride Lipase staining in a formalin-fixed, paraffin-embedded human adipose tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval (EDTA based pH 9.0 solution, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab207799, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. As a negative control (inset), an identical assay was performed without adding the primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Immunohistochemical analysis of paraffin-embedded mouse brown adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse brown adipose tissue is observed (PMID : 15550674). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Immunohistochemical analysis of paraffin-embedded rat white adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat white adipose tissue is observed (PMID : 15550674). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 3T3-L1 (mouse embryonic fibroblast cell line) undifferentiated and differentiated cells labeling Adipose Triglyceride Lipase with ab207799 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing positive staining on 3T3-L1 cells differentiated for 6 days. The level of expression in 3T3/L1 can be induced by differentiation treatment according to the literature (PMID 19297333).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Immunohistochemical analysis of paraffin-embedded mouse white adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse white adipose tissue is observed (PMID : 15550674). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Immunohistochemical analysis of paraffin-embedded rat brown adipose tissue labeling Adipose Triglyceride Lipase with ab207799 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat brown adipose tissue is observed (PMID : 15550674). Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Adipose Triglyceride Lipase was immunoprecipitated from 0.35 mg of 3T3-L1 (mouse embryonic fibroblast cell line) differentiated for 6 days whole cell lysate with ab207799 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207799 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : 3T3-L1 differentiated for 6 days whole cell lysate 10 μg (Input).

Lane 2 : ab207799 IP in 3T3-L1 differentiated for 6 days whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207799 in 3T3-L1 differentiated for 6 days whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/500 dilution

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 1/1000 dilution

Predicted band size: 55 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Western blot : Anti-PNPLA2 antibody [EPR19650] (ab207799) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab207799 was shown to bind specifically to PNPLA2. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in PNPLA2 knockout cell line. To generate this image, wild-type and PNPLA2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PNPLA2 knockout A549 cell lysate at 20 µg

Lane 3:

LNCaP cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin (lane 1) and 3% Milk (lane 2) before being incubated with ab207799 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution

All lanes:

Human adipose normal tissue lysate - total protein (<a href='/en-us/products/unavailable/human-adipose-normal-tissue-lysate-total-protein-ab28980'>ab28980</a>) at 10 µg

Secondary

All lanes:

Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

true

Exposure time: 8min

• WB

Lab

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab207799 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution

Lane 1:

Adult Mouse Adipose Tissue Lysate at 10 µg

Lane 2:

Adult Rat Adipose Tissue Lysate at 10 µg

Secondary

All lanes:

Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

true

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (AB207799)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

Gel type : 4-20% gradient gel (SDS-PAGE)

All lanes:

Western blot - Anti-Adipose Triglyceride Lipase antibody [EPR19650] (ab207799) at 1/1000 dilution

Lane 1:

GST tagged Recombinant Human Adipose Triglyceride Lipase (PNPLA2) protein (Full length, 82 KDa)

Lane 2:

GST tagged Recombinant Human KLF4 protein (Full length, 81 KDa)

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

Predicted band size: 55 kDa

Observed band size: 82 kDa

false

Exposure time: 5s