PKC iota / lambda / zeta in Human Pancreas. PKC iota / lambda / zeta was detected in immersion fixed paraffin-embedded sections of human pancreas using Sheep Anti-Human/Mouse/Rat PKC iota / lambda / zeta Cross-reactive Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4465) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.This application has not been tested in mouse or rat samples.
Detection of Human, Mouse, and Rat PKC iota / lambda / zeta by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line, DU145 human prostate carcinoma cell line, Balb/3T3 mouse embryonic fibroblast cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 1 µg/mL Sheep Anti-Human/Mouse/Rat PKC iota / lambda / zeta Cross-reactive Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4465) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). For additional reference, recombinant human PKC iota, PKC lambda, and PKC zeta (2 ng/lane) were included. Specific bands were detected at approximately 22 kDa for recombinant PKC iota / lambda and ~80 kDa for natural PKC iota / lambda (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
Background: PKC iota/lambda/zeta
Members of the Protein Kinase C (PKC) family are serine/threonine protein kinases that play a key regulatory role in a number of cellular functions including cell growth and differentiation, hormone secretion, and gene expression. Multiple genes and alternative splicing result in three subfamilies, which differ in their co‑factor requirements: conventional PKC isoforms ( alpha, beta Ι, beta ΙΙ, and gamma ) which require calcium and phosphatidylserine (PS), diacylglycerol (DAG) or phorbol esters for activation; novel isoforms (δ, epsilon, eta, and theta ), which are calcium-independent but are still regulated by PS, DAG, or phorbol esters; and atypical isoforms ( iota, lambda, and zeta ), which are calcium-independent and do not require PS, DAG, or phorbol esters for activation. PKC iota has 72% overall identity to PKC zeta. Atypical PKCs have been shown to serve as a convergent downstream target for the PI 3-kinase and TC10 signaling pathways. Stimulation of atypical PKCs by TNF-alpha has been shown to be required for NF‑ kappa B activation. Furthermore, insulin-stimulated atypical PKC activation has been directly implicated in the translocation of GLUT4 and glucose uptake in adipocytes.
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