Anti-PKC delta antibody [14/PKC delta]

4 product images

• 

  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

  Developed using the ECL technique.

  Observed band size: 100 kDa

  Exposure time: 180s

• 

  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] (Anti-GAPDH antibody [EPR16891] - Loading Control 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: 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

  Performed under reducing conditions.

  Observed band size: 75 kDa

• 

  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

  Exposure time: 180s

• 

  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] (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-RABbit IgG H&L (IRDye® 680RD) (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed 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

  Performed under reducing conditions.

  Observed band size: 77 kDa