Anti-PKC delta antibody [14/PKC delta]

• WB

Supplier Data

Western blot - Anti-PKC delta antibody [14/PKC delta] (AB307233)

Blocking/diluting buffer and concentration : 5% NFDM/TBST. The identity of the bands between 100-160 kDa and beneath 40kDa are unknown. Exposure times : Lanes 1, 3-8 : 180 seconds; Lane 2 : 48 seconds.

All lanes:

Western blot - Anti-PKC delta antibody [14/PKC delta] (ab307233) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate 20 μg

Lane 2:

U937 (human histiocytic lymphoma monocyte), whole cell lysate 20 μg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate 20 μg

Lane 4:

Human cerebellum tissue lysate 20 μg

Lane 5:

Mouse cerebellum tissue lysate 20 μg

Lane 6:

Rat cerebellum tissue lysate 20 μg

Lane 7:

C6 (rat glial tumor glial cell), whole cell lysate 10 μg

Lane 8:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate 10 μg

Lane 9:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate 10 μg

Observed band size: 77 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PKC delta antibody [14/PKC delta] (AB307233)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-PKC delta antibody [14/PKC delta] (ab307233) at 1/1000 dilution

Lane 1:

Active human PKC beta 1 fl length protein at 5 ng

Lane 2:

Active human PKC delta fl length protein ng at 5 ng

Lane 3:

Active human PKC zeta fl length protein at 5 ng

Lane 4:

Active human PKC gamma fl length protein at 10 ng

Lane 5:

Active human PKC alpha fl length protein at 60 ng

Lane 6:

Active human PKC beta 2 fl length protein at 5 ng

Lane 7:

Active human PKC eta fl length protein at 5 ng

Lane 8:

Active human PKC epsilon fl length protein at 50 ng

Lane 9:

Active human PKC theta fl length protein at 5 ng

Lane 10:

Active human PKC mu fl length protein at 50 ng

Lane 11:

Active human PKC iota fl length protein at 10 ng

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 100 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PKC delta antibody [14/PKC delta] (AB307233)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS. False colour image of Western blot : Anti-PKC delta antibody [14/PKC delta] (ab307233) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab307233 was shown to bind specifically to 14/PKC delta. A band was observed at 77 kDa in wild-type HeLa cell lysates with no signal observed at this size in the 14/PKC delta knockout cell line. To generate this image, wild-type and 14/PKC delta knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in equal volume of 0.1% TBS 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-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-RABbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. Performed under reducing conditions. The identity of the bands between 100-160 kDa and beneath 40kDa are unknown.

All lanes:

Western blot - Anti-PKC delta antibody [14/PKC delta] (ab307233) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 20 μg

Lane 2:

PRKCD knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 20 μg

Observed band size: 77 kDa

false

• WB

Lab

Western blot - Anti-PKC delta antibody [14/PKC delta] (AB307233)

Western blot : Anti-PRKCD antibody [14/PKC delta] (ab307233) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab307233 was shown to bind specifically to PRKCD. A band was observed at 75 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in PRKCD knockout cell line. To generate this image, wild-type and PRKCD knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PKC delta antibody [14/PKC delta] (ab307233) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

Western blot - Human PRKCD knockout U-87 MG cell line (<a href='/en-us/products/cell-lines/human-prkcd-knockout-u-87-mg-cell-line-ab306756'>ab306756</a>)

Lane 2:

PRKCD knockout U-87 MG cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 75 kDa

false