Anti-PKC epsilon antibody [EPR28194-50] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-PKC epsilon antibody [EPR28194-50] - BSA and Azide free (AB317499)

This data was developed using ab317498, the same antibody clone in a different buffer formulation.

The identity of the lower MW band at approximately 26 kDa (in lanes 1-7) are unknown.

The identity of the higher MW band at approximately 240 kDa (only detected in fresh cells) is unknown.

In lanes 1-3, the lysate was stored at -80°C prior to Western Blotting. The bands between 37 kDa and 84 kDa are likely to be degradation products. In lanes 4-7, 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.

All lanes:

Western blot - Anti-PKC epsilon antibody [EPR28194-50] (<a href='/en-us/products/primary-antibodies/pkc-epsilon-antibody-epr28194-50-ab317498'>ab317498</a>) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lanes 2 and 6:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lanes 3 and 7:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 85 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PKC epsilon antibody [EPR28194-50] - BSA and Azide free (AB317499)

This data was developed using ab317498, the same antibody clone in a different buffer formulation.

Low expression : HEL, HepG2.

The identity of the lower MW band at approximately 26 kDa (in lanes 1-3) is unknown.

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

All lanes:

Western blot - Anti-PKC epsilon antibody [EPR28194-50] (<a href='/en-us/products/primary-antibodies/pkc-epsilon-antibody-epr28194-50-ab317498'>ab317498</a>) at 1/1000 dilution

Lane 1:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEL (human erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) 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: 85 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PKC epsilon antibody [EPR28194-50] - BSA and Azide free (AB317499)

This data was developed using ab317498, the same antibody clone in a different buffer formulation.

Low expression : heart (PMID : 1556149), spleen (PMID : 1556149), skeletal muscle.

The identity of the bands between 25 kDa and 50 kDa are unknown.

Lanes 6-11 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 1-5 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

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

All lanes:

Western blot - Anti-PKC epsilon antibody [EPR28194-50] (<a href='/en-us/products/primary-antibodies/pkc-epsilon-antibody-epr28194-50-ab317498'>ab317498</a>) at 1/1000 dilution

Lane 1:

Human hypothalamus tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

Human hippocampus tissue lysate at 20 µg

Lane 4:

Human skeletal muscle tissue lysate at 20 µg

Lane 5:

Human spleen tissue lysate at 20 µg

Lane 6:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 7:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 8:

Mouse heart tissue lysate at 20 µg

Lane 9:

Rat cerebral cortex tissue lysate at 20 µg

Lane 10:

Rat skeletal muscle tissue lysate at 20 µg

Lane 11:

Rat heart tissue lysate at 20 µg

Secondary

Lanes 1 - 5:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 6 - 11:

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: 85 kDa,36 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-PKC epsilon antibody [EPR28194-50] - BSA and Azide free (AB317499)

This data was developed using ab317498, the same antibody clone in a different buffer formulation.

This antibody does not cross-react with human PKC alpha, PKC beta 1, PKC beta 2, PKC gamma, PKC delta, PKC eta, PKC theta, PKC mu.

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

All lanes:

Western blot - Anti-PKC epsilon antibody [EPR28194-50] (<a href='/en-us/products/primary-antibodies/pkc-epsilon-antibody-epr28194-50-ab317498'>ab317498</a>) at 1/1000 dilution

Lane 1:

His-tagged Human PKC epsilon fragment at 10 ng

Lane 2:

GST-tagged full-length Human PKC alpha recombinant protein at 10 ng

Lane 3:

GST-tagged full-length Human PKC beta 1 recombinant protein at 10 ng

Lane 4:

GST-tagged full-length Human PKC beta 2 recombinant protein at 10 ng

Lane 5:

GST-tagged full-length Human PKC gamma recombinant protein at 10 ng

Lane 6:

GST-tagged full-length Human PKC delta recombinant protein at 10 ng

Lane 7:

GST-tagged full-length Human PKC eta recombinant protein at 10 ng

Lane 8:

GST-tagged full-length Human PKC theta recombinant protein at 10 ng

Lane 9:

GST-tagged full-length Human PKC mu 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 1/100000 dilution

Observed band size: 26 kDa,85 kDa

false

Exposure time: 180s