Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (AB317610)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 3T3-L1 (mouse embryonic fibroblast) cells labelling Adipose Triglyceride Lipase with ab317610 at 1/50 (10.08 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 cytoplasmic and membranous staining in differentiated 3T3-L1 cells (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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). 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-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (AB317610)

Adipose Triglyceride Lipase was immunoprecipitated from 0.35 mg Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days whole cell lysate with ab317610 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317610 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days whole cell lysate
Lane 2 : ab317610 IP in Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317610 in differentiated 3T3-L1 for 6 days whole cell lysate

All lanes:

Immunoprecipitation - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (ab317610) at 1/30 dilution

All lanes:

Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days whole cell lysate at 10 µg

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

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Exposure time: 180s

• WB

Lab

Western blot - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (AB317610)

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

Exposure time :
Lane 1-2 : 5 seconds;
Lane 3-4 : 180 seconds

ab181602 was used as a GAPDH loading control at 1/200000 dilution

All lanes:

Western blot - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (ab317610) at 1/1000 dilution

Lane 1:

Human colon tissue lysate (untreated membrane) at 20 µg

Lane 2:

Human colon tissue lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 3:

Human fat tissue lysate (untreated membrane) at 20 µg

Lane 4:

Human fat tissue lysate (Alkaline phosphatase treated membrane) 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 1/20000 dilution

Observed band size: 55 kDa

false

• WB

Supplier Data

Western blot - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (AB317610)

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

All lanes:

Western blot - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (ab317610) at 1/1000 dilution

Lane 1:

Undifferentiated 3T3-L1 (mouse embryonic fibroblast) whole cell lysate (untreated membrane) at 40 µg

Lane 2:

3T3-L1 differentiated for 6 days whole cell lysate (untreated membrane) at 40 µg

Lane 3:

Undifferentiated 3T3-L1 whole cell lysate (Lambda phosphatase treated membrane) at 40 µg

Lane 4:

3T3-L1 differentiated for 6 days whole cell lysate (Lambda phosphatase treated membrane) at 40 µg

Lane 5:

Mouse white fat tissue lysate (untreated membrane) at 40 µg

Lane 6:

Mouse white fat tissue lysate (Lambda phosphatase treated membrane) at 40 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: 55 kDa,36 kDa

false

Exposure time: 180s

• ELISA

Supplier Data

ELISA - Anti-Adipose Triglyceride Lipase (phospho S406) antibody [EPR27159-29] (AB317610)

Indirect ELISA analysis of ab317610 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Mouse Adipose Triglyceride Lipase(phospho S406),Mouse Adipose Triglyceride Lipase non-phospho,Mouse Adipose Triglyceride Lipase(mutated S406A), Human Adipose Triglyceride Lipase(phospho S404),Human Adipose Triglyceride Lipase non-phospho.

Antigen concentration : 100ng/ml