Anti-PKC eta antibody [EPR18513]

• IP

Supplier Data

Immunoprecipitation - Anti-PKC eta antibody [EPR18513] (AB179524)

PKC eta was immunoprecipitated from 1mg of Mouse lung whole cell lysate with ab179524 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab179524 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : Mouse lung whole cell lysate 10ug (Input).

Lane 2 : ab179524 IP in Mouse lung whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab179524 in Mouse lung whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-PKC eta antibody [EPR18513] (ab179524)

Predicted band size: 78 kDa

Observed band size: 40 kDa,78 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Blocking/Dilution buffer : 5% NFDM/TBST.

The antibody recognizes the full length (~78KD) and the catalytic domain (~40KD) of PKC eta.
(Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331; Newton AC. 1995. J. Biol. Chem. 270, 28495-28498)

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (ab179524) at 1/10000 dilution

Lane 1:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 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: 78 kDa

Observed band size: 45 kDa,78 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Blocking/Dilution buffer : 5% NFDM/TBST.

The antibody recognizes the full length (~78KD) and the catalytic domain (~40KD) of PKC eta.
(Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331; Newton AC, 1995. J. Biol. Chem. 270, 28495-28498)

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (ab179524) at 1/2000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 78 kDa

Observed band size: 45 kDa,78 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Blocking/Dilution buffer : 5% NFDM/TBST.

The antibody recognizes the full length (~78KD) and the catalytic domain (~40KD) of PKC eta.
(Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331; Newton AC, 1995. J. Biol. Chem. 270, 28495-28498)

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (ab179524) at 1/10000 dilution

Lane 1:

Human fetal skin lysate at 10 µg

Lane 2:

Mouse lung lysate at 10 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: 78 kDa

Observed band size: 45 kDa,78 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Blocking/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 eta antibody [EPR18513] (ab179524) 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: 78 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Western blot : Rabbit Monoclonal [EPR18513] to PKC ab179524 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 76, 85 kDa in Wild-type A549 cell lysates with no signal observed at this size in PRKCH knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^™ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (ab179524) at 1/2000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

PRKCH knockout A549 whole cell lysate at 20 µg

Lane 3:

U-2 OS whole cell lysate at 20 µg

Lane 4:

HL-60 whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 76 kDa,85 kDa

false

• WB

Supplier Data

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Blocking/Dilution buffer : 5% NFDM/TBST.

The antibody recognizes the full length (~78KD) and the catalytic domain (~40KD) of PKC eta.
(Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331; Newton AC, 1995. J. Biol. Chem. 270, 28495-28498)

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (ab179524) at 1/2000 dilution

Lane 1:

Rat skin lysate at 10 µg

Lane 2:

Mouse skin lysate at 10 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: 78 kDa

Observed band size: 45 kDa,78 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PKC eta antibody [EPR18513] (AB179524)

Blocking/Dilution buffer : 5% NFDM/TBST.

The antibody recognizes the full length (~78KD) and the catalytic domain (~40KD) of PKC eta.
(Geraldes P and King GL, 2010. Circ Res. 106, 1319-1331; Newton AC, 1995. J. Biol. Chem. 270, 28495-28498)

All lanes:

Western blot - Anti-PKC eta antibody [EPR18513] (ab179524) at 1/10000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Rat brain lysate at 10 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: 78 kDa

Observed band size: 45 kDa,78 kDa

false

Exposure time: 30s