Anti-Perilipin 3/TIP47 antibody [EPR28801-93]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (AB315476)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Perilipin 3/TIP47 with ab315476 at 1/1000 (0.518 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 staining in HeLa cell line. 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 (Red). 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (AB315476)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Perilipin 3/TIP47 with ab315476 at 1/50 dilution (1 ug)/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.

• IP

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Immunoprecipitation - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (AB315476)

Perilipin 3/TIP47 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab315476 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315476 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab315476 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315476 in HeLa whole cell lysate

The bands beneath the target band (47 kDa) are likely to be degraded target fragments in lane 2.

All lanes:

Immunoprecipitation - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (ab315476) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• WB

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Western blot - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (AB315476)

The bands beneath the target band (47 kDa) are likely 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-Perilipin 3/TIP47 antibody [EPR28801-93] (ab315476) at 1/1000 dilution

All lanes:

Human placenta 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: 47 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (AB315476)

In lanes 1-4, the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (47 kDa) are likely to be degradation products. In lanes 5-7, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

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

Exposure time : Lanes 1-6 : 180 seconds, lane 7 : 26 seconds.

All lanes:

Western blot - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (ab315476) at 1/1000 dilution

Lanes 1 and 5:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 2 and 7:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

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

Lane 6:

293T (human embryonic kidney epithelial cell) 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: 47 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (AB315476)

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

All lanes:

Western blot - Anti-Perilipin 3/TIP47 antibody [EPR28801-93] (ab315476) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting Perilipin 3 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: 47 kDa,36 kDa

false

Exposure time: 180s