Anti-PKC antibody [EPR16898] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PKC alpha + beta + gamma with ab179522 at 1/200 dilution, followed by Goat anti-rabbit IAlexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image shows cytoplasmic staining on K562 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor®594 secondary antibody) at 1/500 dilution (red).

The negative controls are as follows : -ve control 1 : ab179522 at 1/200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed NIH/3T3 (Mouse embyro fibroblast cells) cells labeling PKC alpha + beta + gamma with ab179522 at 1/360 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

PKC alpha + beta + gamma was immunoprecipitated from 1mg of Rat brain extract with ab179522 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab179522 at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody was used as secondary antibody at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>)

Predicted band size: 77 kDa

false

• WB

Lab

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Active human PKC zeta full length protein (ab60848) contains aa1-592 with GST-tag; Active human PKC iota full length protein (ab60850) contains aa1-596 with GST-tag.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/2000 dilution

Lane 1:

Active human PKC zeta full length protein at 0.02 µg

Lane 2:

Active human PKC iota full length protein at 0.02 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 1/50000 dilution

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/1000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

Human fetal heart tissue lysate at 10 µg

Lane 3:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/10000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

293T (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Lane 3:

SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Lab

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Active human PKC alpha full length protein (ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein (ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein (ab60841) contains aa1-673 with GST-tag; Active human PKC gamma full length protein (ab60842) contains aa1-677 with GST-tag; Active human PKC delta full length protein (ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein (ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein (ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein (ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein (ab60873) contains aa1-912 with GST-tag.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/2000 dilution

Lane 1:

Active human PKC alpha full length protein at 0.02 µg

Lane 2:

Active human PKC beta 1 full length protein at 0.02 µg

Lane 3:

Active human PKC beta 2 full length protein at 0.02 µg

Lane 4:

Active human PKC gamma full length protein at 0.02 µg

Lane 5:

Active human PKC delta full length protein at 0.02 µg

Lane 6:

Active human PKC eta full length protein at 0.02 µg

Lane 7:

Active human PKC epsilon full length protein at 0.02 µg

Lane 8:

Active human PKC theta full length protein at 0.02 µg

Lane 9:

Active human PKC mu full length protein at 0.02 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 1/50000 dilution

Predicted band size: 77 kDa

false

Exposure time: 2s

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/1000 dilution

Lane 1:

K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate at 20 µg

Lane 2:

A431 (Human epidermoid carcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/5000 dilution

Lane 1:

L6 (Rat skeletal muscle cell line) whole cell lysate at 10 µg

Lane 2:

Neuro-2a (Mouse neuroblastoma cells) whole cell lysate at 10 µg

Lane 3:

L-929 (Mouse connective tissue fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>) at 1/1000 dilution

All lanes:

Rat brain whole tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 77 kDa

Observed band size: 77 kDa

false

• WB

Unknown

Western blot - Anti-PKC antibody [EPR16898] - BSA and Azide free (AB250108)

This data was developed using ab179522, the same antibody clone in a different buffer formulation.

Recombinant PKC isoforms were tested for reactivity by western blot, and are marked as positive (+) or negative (-).

All lanes:

Western blot - Anti-PKC antibody [EPR16898] (<a href='/en-us/products/primary-antibodies/pkc-antibody-epr16898-ab179522'>ab179522</a>)

false