Invitrogen

PKC zeta Polyclonal Antibody

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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PKC zeta Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of PKC zeta was performed using 70% confluent log phase HeLa and HL-60 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with PKC zeta Polyclonal Antibody (Product # PA5-78129) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing endosome pattern of staining. Panel e shows HL-60 cells with no expression of PKC zeta. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PKC zeta Antibody in Western Blot (WB) — Western blot was performed using Anti-PKC zeta Polyclonal Antibody (Product # PA5-78129) and a 67kDa band corresponding to PKC zeta was observed in all cell lines and tissue lysates tested except HL-60 and Mouse Skeletal Muscle which are reported to be negative. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), MDA-MB-231 (Lane 4), THP-1 (Lane 5), HL-60 (Lane 6), Mouse Brain (Lane 7) and Mouse Skeletal Muscle (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

PKC zeta Antibody in Immunoprecipitation (IP) — Immunoprecipitation of PKC zeta was performed in 293T whole cell extracts using 5 µg of PKC zeta Polyclonal Antibody (Product # PA5-78129). Samples were transferred to a membrane and probed with PKC zeta Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PKC zeta Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Immunofluorescence analysis of PKC zeta shows increased expression of proteins in HeLa cell line when compared to HL-60 using Anti-PKC zeta Polyclonal Antibody (Product # PA5-78129). {RE}

Product Details

PA5-78129

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	-
Immunocytochemistry (ICC/IF)	1:100	-
Immunoprecipitation (IP)	1:100-1:500	-

Product Specifications
Species Reactivity	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Carrier-protein conjugated synthetic peptide encompassing a sequence within the C-terminus region of human PKC zeta. The exact sequence is proprietary. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 1% BSA, 20% glycerol
Contains	0.025% ProClin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2736440

Product Specific Information

Positive Control: 293T, A431, HeLa, HepG2

Predicted Reactivity: Mouse (100%), Rat (100%), Zebrafish (100%), Pig (100%), Rabbit (100%), Bovine (100%)

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

PKC zeta is a member of the protein kinase C (PKC) family play a key regulatory role in a variety of cellular functions including cell growth and differentiation, gene expression, hormone secretion and membrane function. PKCs were originally identified as serine/threonine protein kinases whose activity was dependent on calcium and phospholipids. Diacylglycerols (DAG) and tumor promoting phorbol esters bind to and activate PKC. PKCs can be subdivided into at least two major classes including conventional (c) PKC isoforms ( alpha, betaI, betaII and gamma) and novel (n) PKC isoforms ( delta, epsilon, zeta, eta and theta). Patterns of expression for each PKC isoform differ among tissues and PKC family member exhibit clear differences in their cofactor dependencies. For instance, the kinase activities of nPKC delta and epsilon are independent of Ca2+.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: atypical protein kinase C; kinase nPKC-zeta; KPCZ; nPKC-zeta; nPKC-zeta PRKCZ; protein kinase C zeta subspecies; Protein kinase C zeta type; protein kinase C, zeta

Gene Aliases: AI098070; aPKCzeta; C80388; nPKC-zeta; PKC-ZETA; PKC2; Pkcz; PRKCZ; R74924; zetaPKC

UniProt ID: (Human) Q05513, (Mouse) Q02956

Entrez Gene ID: (Human) 5590, (Mouse) 18762

Molecular Function: non-receptor serine/threonine protein kinase