Anti-PDE3B antibody [EPR28229-71] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-PDE3B antibody [EPR28229-71] - BSA and Azide free (AB317027)

This data was developed using ab317026, the same antibody clone in a different buffer formulation.

PDE3B was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab317026 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317026 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 : ab317026 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317026 in HeLa whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-PDE3B antibody [EPR28229-71] (<a href='/en-us/products/primary-antibodies/pde3b-antibody-epr28229-71-ab317026'>ab317026</a>) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

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: 24s

• WB

Supplier Data

Western blot - Anti-PDE3B antibody [EPR28229-71] - BSA and Azide free (AB317027)

This data was developed using ab317026, the same antibody clone in a different buffer formulation.

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

The identity of the lower MW band at approximately 20 kDa is unknown.

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

All lanes:

Western blot - Anti-PDE3B antibody [EPR28229-71] (<a href='/en-us/products/primary-antibodies/pde3b-antibody-epr28229-71-ab317026'>ab317026</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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: 145 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PDE3B antibody [EPR28229-71] - BSA and Azide free (AB317027)

This data was developed using ab317026, the same antibody clone in a different buffer formulation.

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

The identity of the lower MW band at approximately 20 kDa is unknown.

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

All lanes:

Western blot - Anti-PDE3B antibody [EPR28229-71] (<a href='/en-us/products/primary-antibodies/pde3b-antibody-epr28229-71-ab317026'>ab317026</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

HeLa transfected with siRNA specifically targeting PDE3B whole cell lysate at 20 µg with 5% NFDM/TBST

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: 145 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-PDE3B antibody [EPR28229-71] - BSA and Azide free (AB317027)

This data was developed using ab317026, the same antibody clone in a different buffer formulation.

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

Low expression : human skeletal muscle.

The identity of the lower MW band at approximately 20 kDa is unknown.

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

All lanes:

Western blot - Anti-PDE3B antibody [EPR28229-71] (<a href='/en-us/products/primary-antibodies/pde3b-antibody-epr28229-71-ab317026'>ab317026</a>) at 1/1000 dilution

Lane 1:

Human adipocytes tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human 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: 145 kDa,36 kDa

false

Exposure time: 70s

• Dot

Supplier Data

Dot Blot - Anti-PDE3B antibody [EPR28229-71] - BSA and Azide free (AB317027)

This data was developed using ab317026, the same antibody clone in a different buffer formulation.

Dot blot analysis of PDE3B using ab317026 at 1 : 1000 (0.498 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

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

This antibody does not cross-react with human PDE3A.

All lanes:

Dot Blot - Anti-PDE3B antibody [EPR28229-71] (<a href='/en-us/products/primary-antibodies/pde3b-antibody-epr28229-71-ab317026'>ab317026</a>) at 1/1000 dilution

Lane 1:

His-tagged human PDE3B protein

Lane 2:

His-tagged human PDE3A protein

Secondary

All lanes:

Dot 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

false

Exposure time: 180s