ab59364 detects endogenous levels of total PKC zeta protein. The immunogen used for this product shares 89% homology with PKC iota/lambda. Cross-reactivity with this protein has not been confirmed experimentally.
Synthetic peptide within Human PKC zeta aa 526-575. The exact sequence is proprietary. Synthesized non-phosphopeptide derived from human PKC zeta around the phosphorylation site of threonine 560 (Q-L-TP-P-D). Database link: Q05513
WB: Xenopus laevis cytoplasmic egg extract. PMA treated COS7 cell extracts. Rat hippocampus lysate.
IP: PKC zeta IP in mouse brain tissue lysate.
ICC/IF: NIH/3T3 cells.
IHC-P: Human proliferative endometrial tissue. Human brain tissue.
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This is a calcium-independent, phospholipid-dependent, serine- and threonine-specific enzyme. Phosphorylates 'Ser-311' of RELA subunit of NF-kappa-B. PKC is activated by diacylglycerol which in turn phosphorylates a range of cellular proteins. PKC also serves as the receptor for phorbol esters, a class of tumor promoters. Subunit of a quaternary complex that plays a central role in epithelial cell polarization.
Expressed in brain, and to a lesser extent in lung, kidney and testis.
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily. Contains 1 AGC-kinase C-terminal domain. Contains 1 OPR domain. Contains 1 phorbol-ester/DAG-type zinc finger. Contains 1 protein kinase domain.
The OPR domain mediates mutually exclusive interactions with SQSTM1 and PARD6B. The C1 domain does not bind the diacylglycerol (DAG).
CDH5 is required for its phosphorylation at Thr-410.
Cytoplasm. Endosome. Cell junction. In the retina, localizes in the terminals of the rod bipolar cells (By similarity). Associates with endosomes. Presence of KRIT1, CDH5 and RAP1B is required for its localization to the cell junction.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC zeta antibody (ab59364)Image from Rivero R et al., Reproductive Biology and Endocrinology 2012, 10:17, 5 March 2012. Fig 1.; doi: 10.1186/1477-7827-10-17.
Immunohistochemical analysis of human proliferative endometrial tissue, staining PKC zeta with ab59364.
Antigen retrieval was performed by heat mediation in a citrate buffer. Tissue was blocked with blocking solution before incubating with primary antibody (1/100) overnight at 4°C. Staining was detected using DAB.
PKC zeta was immunoprecipitated from 0.5 mg mouse brain tissue with 5 µg of ab59364 and 50 µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10 minutes; Mouse brain tissue lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 minutes under agitation.
Proteins were eluted by addition of 40 µl SDS loading buffer and incubated for 10 minutes at 70°C; 10 µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab59364.
Band: 68kDa; PKC zeta, non specific bands - 52kDa: We are unsure as to the identity of this extra band.
Western blot - Anti-PKC zeta antibody (ab59364)
All lanes : Anti-PKC zeta antibody (ab59364) at 1/500 dilution
Lane 1 : Extracts from COS-7 (African green monkey kidney fibroblast-like cell line) cells, treated with PMA (125 ng/ml, 30mins), without immunizing peptide Lane 2 : Extracts from COS7 (African green monkey kidney fibroblast-like cell line)cells, treated with PMA (125 ng/ml, 30mins), plus immunizing peptide