Anti-Perilipin-1 antibody [EPR28189-66] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Perilipin-1 antibody [EPR28189-66] - BSA and Azide free (AB317261)

This data was developed using ab317260, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 3T3-L1 (mouse embryonic fibroblast) cells labelling Perilipin-1 with ab317260 at 1/1000 (0.493 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 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-Perilipin-1 antibody [EPR28189-66] - BSA and Azide free (AB317261)

This data was developed using ab317260, the same antibody clone in a different buffer formulation.

Perilipin-1 was immunoprecipitated from 0.35 mg Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days whole cell lysate with ab317260 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317260 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 : ab317260 IP in Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317260 in differentiated 3T3-L1 for 6 days whole cell lysate.

The bands beneath the target band (55 kDa) are likely to be degraded target fragments (PMID : 16448845).

All lanes:

Immunoprecipitation - Anti-Perilipin-1 antibody [EPR28189-66] (<a href='/en-us/products/primary-antibodies/perilipin-1-antibody-epr28189-66-ab317260'>ab317260</a>) at 1/30 dilution

All lanes:

Differentiated 3T3-L1(mouse embryonic fibroblast) for 6 days 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: 10s

• WB

Supplier Data

Western blot - Anti-Perilipin-1 antibody [EPR28189-66] - BSA and Azide free (AB317261)

This data was developed using ab317260, the same antibody clone in a different buffer formulation.

Low expression : mouse skeletal muscle (PMID : 32996002).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

The bands beneath the target band (55 kDa) are likely to be degraded target fragments (PMID : 16448845).

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

Exposure time : Lane 1-2 : 37 seconds; Lane 3 : 180 seconds

All lanes:

Western blot - Anti-Perilipin-1 antibody [EPR28189-66] (<a href='/en-us/products/primary-antibodies/perilipin-1-antibody-epr28189-66-ab317260'>ab317260</a>) at 1/1000 dilution

Lane 1:

Mouse white fat tissue lysate at 20 µg

Lane 2:

Mouse brown fat tissue lysate at 20 µg

Lane 3:

Mouse 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 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Perilipin-1 antibody [EPR28189-66] - BSA and Azide free (AB317261)

This data was developed using ab317260, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-Perilipin-1 antibody [EPR28189-66] (<a href='/en-us/products/primary-antibodies/perilipin-1-antibody-epr28189-66-ab317260'>ab317260</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

Differentiated 3T3-L1 for 6 days 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 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 180s