Overview

  • Product name
  • Description
    Rabbit polyclonal to PKC zeta
  • Specificity
    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.
  • Tested applications
    Suitable for: WB, ELISA, IHC-P, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis
  • Immunogen

    Synthesized non-phosphopeptide derived from human PKC zeta around the phosphorylation site of threonine 560 (Q-L-TP-P-D).

  • Positive control
    • COS7 cytoplasmic lysate Human brain tissue

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • Purification notes
    Affinity-purified from rabbit antiserum by affinity-chromatography, using epitope-specific immunogen.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab59364 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500 - 1/1000. Predicted molecular weight: 68 kDa.
ELISA 1/1000.
IHC-P 1/50 - 1/100.
IP Use a concentration of 5 µg/ml.
ICC/IF 1/100 - 1/500.

Target

  • Function
    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.
  • Tissue specificity
    Expressed in brain, and to a lesser extent in lung, kidney and testis.
  • Sequence similarities
    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.
  • Domain
    The OPR domain mediates mutually exclusive interactions with SQSTM1 and PARD6B.
    The C1 domain does not bind the diacylglycerol (DAG).
  • Post-translational
    modifications
    CDH5 is required for its phosphorylation at Thr-410.
  • Cellular localization
    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.
  • Information by UniProt
  • Database links
  • Alternative names
    • 14-3-3-zetaisoform antibody
    • AI098070 antibody
    • aPKCzeta antibody
    • C80388 antibody
    • EC 2.7.11.13 antibody
    • KPCZ_HUMAN antibody
    • nPKC zeta antibody
    • nPKC-zeta antibody
    • OTTHUMP00000001368 antibody
    • OTTHUMP00000044160 antibody
    • PKC 2 antibody
    • PKC ZETA antibody
    • PKC2 antibody
    • Pkcz antibody
    • PKCZETA antibody
    • PKM-zeta, included antibody
    • PRKCZ antibody
    • Protein kinase C zeta antibody
    • Protein kinase C zeta form antibody
    • Protein kinase C zeta type antibody
    • r14-3-3 antibody
    • R74924 antibody
    • zetaPKC antibody
    see all

Images

  • PKC zeta was immunoprecipitated using 0.5mg Mouse Brain tissue, 5µg of Rabbit polyclonal to PKC zeta and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain tissue lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab59364.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Anti-Rabbit IgG light chain HRP (ab99697).

    Band: 68kDa; PKC zeta, non specific bands - 52kDa: We are unsure as to the identity of this extra band.

  • All lanes : Anti-PKC zeta antibody (ab59364) at 1/500 dilution

    Lane 1 : Extracts from COS7 cells, treated with PMA (125ng/ml, 30mins), minus immunising peptide
    Lane 2 : Extracts from COS7 cells, treated with PMA (125ng/ml, 30mins), plus immunising peptide


    Predicted band size : 68 kDa
    Observed band size : 80 kDa (why is the actual band size different from the predicted?)
  • Lane 1 :
    Lanes 2 - 4 : Anti-PKC zeta antibody (ab59364) at 1/500 dilution

    Lane 1 : 40 µg Hippocampus lysate
    Lane 2 : 40 µg Hippocampus lysate
    Lane 3 : 20 µg Hippocampus lysate
    Lane 4 : 20 µg Hippocampus lysate

    Secondary
    Anti-Rabbit IR800-linked conjugation: IRDye® 800CW

    Performed under reducing conditions.

    Predicted band size : 68 kDa

    This image is courtesy of an Abreview submitted by Mr Richard D'Mello

    Detection method: Odyssey Detection System Exposure: 5 minutes and 0 seconds Bands: Specific: 68 kDa

    See Abreview



  • Predicted band size : 68 kDa
    Lane 2: anti-PKC zeta (ab59364) blot of X. laevis cytoplasmic egg extract, 1:500

    See Abreview

  • Immunofluorescence analysis of NIH/3T3 cells using PKC zeta at dilution of 1:100.

  • 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.
  • ab59364, at a 1/50 dilution, staining humna PKC zeta in brain, using Immunohistochemistry, Formalin/PFA fixed Paraffin embedded tissue, minus immunising peptide (left image) and plus immunising peptide (right image).

References

This product has been referenced in:
  • Dong Z  et al. Long-term potentiation decay and memory loss are mediated by AMPAR endocytosis. J Clin Invest 125:234-47 (2015). Read more (PubMed: 25437879) »
  • Corriden R  et al. Tamoxifen augments the innate immune function of neutrophils through modulation of intracellular ceramide. Nat Commun 6:8369 (2015). Read more (PubMed: 26458291) »

See all 5 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (X. laevis early embryo (4 cell))
Specification
X. laevis early embryo (4 cell)
Fixative
DENTS (80% Methanol, 20% DMSO)
Permeabilization
Yes - PBS 0.1% Triton-X100
Blocking step
PBS 0.1% Triton-X100, 2 mg/ml BSA, 10% goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Xenopus laevis Cell lysate - other (Xenopus laevis cytoplasmic egg extract)
Loading amount
40 µg
Specification
Xenopus laevis cytoplasmic egg extract
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Hippocampus lysate)
Loading amount
40 µg
Specification
Hippocampus lysate
Gel Running Conditions
Reduced Denaturing (8% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Mr. Richard D'Mello

Verified customer

Submitted Apr 08 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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