Anti-PPAR gamma antibody [RM1091]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PPAR gamma antibody [RM1091] (AB316981)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) (Right) / Jurkat(human T cell leukemia T lymphocyte from peripheral blood) (Left) cells labelling PPAR gamma with ab316981 at 1/50 dilution (1ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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

Negative control : Jurkat (PMID : 7787419).
Anti-PPAR gamma antibody (ab316981) is specific to human species, while Anti-PPAR gamma antibody (ab209350) is non-specific.

• IP

Supplier Data

Immunoprecipitation - Anti-PPAR gamma antibody [RM1091] (AB316981)

PPAR gamma was immunoprecipitated from 0.35 mg PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate with ab316981 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316981 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab316981 IP in PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316981 in PC-3 whole cell lysate

All lanes:

Immunoprecipitation - Anti-PPAR gamma antibody [RM1091] (ab316981) at 1/30 dilution

All lanes:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate

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

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PPAR gamma antibody [RM1091] (AB316981)

Negative control : MOLT-4 and Jurkat (PMID : 7787419).

The expression of PPARG is upregulated in response to TPA treatment ( PMID : 9568716/PMID : 19850645).

The bands beneath the target band (53kDa) are expected to be degraded target fragments.

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

All lanes:

Western blot - Anti-PPAR gamma antibody [RM1091] (ab316981) at 1/1000 dilution

Lane 1:

HDLM-2 (human Hodgkin lymphoma cell) whole cell lysate at 20 µg

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 3 - 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 5:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 6:

THP-1 treated with 80nM TPA for 24 hours, whole cell lysate 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: 53 kDa,58 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PPAR gamma antibody [RM1091] (AB316981)

Anti-PPAR gamma antibody (ab316981) staining at 1/1000 dilution is specific to human, mouse and rat species. Anti-PPAR gamma antibody (ab209350) at 1/1000 dilution is bandy than ab316981.

The expression profile observed is consistent with what has been described in the literature (PMID : 9065481).

The bands beneath the target band (53kDa) are expected to be degraded target fragments.

All lanes:

Western blot - Anti-PPAR gamma antibody [RM1091] (ab316981) at 1/1000 dilution

Lane 1:

Human fat tissue lysate at 20 µg

Lane 2:

Mouse brown fat tissue lysate at 20 µg

Lane 3:

Mouse ovary tissue lysate at 20 µg

Lane 4:

Mouse colon cancer tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 6:

Rat brown fat tissue lysate at 20 µg

Lane 7:

Rat ovary tissue lysate at 20 µg

Lane 8:

Rat colon tissue lysate at 20 µg

Lane 9:

Rat skeletal muscle tissue lysate 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: 53 kDa,58 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PPAR gamma antibody [RM1091] (AB316981)

Negative control : NIH/3T3 (PMID : 12023027).

The expression of PPARG is upregulated in response to 3T3-L1 differentiation for 6 days treatment (PMID : 28759152).

The bands beneath the target band (53kDa) are expected to be degraded target fragments.

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

All lanes:

Western blot - Anti-PPAR gamma antibody [RM1091] (ab316981) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

NIH/3T3 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: 53 kDa,58 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PPAR gamma antibody [RM1091] (AB316981)

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-PPAR gamma antibody [RM1091] (ab316981) at 1/1000 dilution

Lane 1:

His-tagged PPAR delta recombinant protein at 10 ng

Lane 2:

His-tagged PPAR alpha recombinant protein at 10 ng

Lane 3:

His-GST-tagged PPAR gamma recombinant protein at 10 ng

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: 85 kDa

true

Exposure time: 180s