Human PKC zeta Antibody

Name: Human PKC zeta Antibody
Brand: R&D Systems
SKU: AF6666
Price: 399 USD
Availability: InStock

R&D Systems | Catalog # AF6666

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
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Specific Notices
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human

Applications

Knockout Validated, Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

View all PKC zeta Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human PKC zeta
Ile454-Val592
Accession # Q05513

Specificity

Detects human PKC zeta in Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human PKC zeta Antibody

Detection of Human PKC zeta by Western Blot.

Western blot shows lysates of 293T human embryonic kidney cell line, MCF-7 human breast cancer cell line, and human lung tissue. PVDF Membrane was probed with 1 µg/mL of Human PKC zeta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6666) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PKC zeta at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human PKC zeta Specificity by Using Knockout Cell Line.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and PKC zeta knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human PKC zeta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6666) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PKC zeta at approximately 75 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human PKC zeta by Simple Western^TM.

Simple Western shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.5 mg/ml. A specific band was detected for PKC zeta at approximately 79 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human PKC zeta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6666). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Applications for Human PKC zeta Antibody

Application

Recommended Usage

Knockout Validated

PKC zeta is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in PKC zeta knockout HEK293T cell line.

Simple Western

10 µg/mL
Sample: MCF-7 human breast cancer cell line

Western Blot

1 µg/mL
Sample: 293T human embryonic kidney cell line, MCF‑7 human breast cancer cell line, and human lung tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: PKC zeta

PKC zeta (protein kinase C-zeta, EC=2.7.11.13), also called PKC2, is a 592 amino acid (aa), 79 kDa member of the atypical PKC (aPKC) subfamily of Ser/Thr protein kinases. aPKC proteins can be activated by phorbol esters, but are insensitive to calcium and diacylglycerol. PKC zeta regulates cell polarity in early development, and is necessary for cytokine-induced chemotaxis of macrophages and cancer cells, chemokine-induced adhesion of neutrophils, and polarization of Th2 cells. It is activated by PDK1 phosphorylation, binds ceramide, and translocates to the plasma membrane where it regulates adherens and tight junction formation. PKC zeta contains an OPR domain (aa 15-98), a zinc finger region (aa 130-180), and a protein kinase domain (aa 252-518). PKC zeta potential variants of 409 or 539 aa have alternate start sites at aa 184 or aa 9 (also missing aa 186-233), respectively. Within the region used as an immunogen, human PKC zeta shares 98% aa identity with mouse and rat PKC zeta.