PKC iota/lambda/zeta Antibody [Unconjugated] - Cross-reactive

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 zeta, and PKC alpha (2 ng/lane) were included. Specific bands were detected at approximately 22 kDa for recombinant PKC iota/zeta and ~80 kDa for natural PKC iota/zeta (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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.

Simple Western shows lysates of A431 human epithelial carcinoma cell line, loaded at 0.5 mg/ml. A specific band was detected for PKC iota / lambda / zeta at approximately 80 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse/Rat PKC iota / lambda / zeta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4465). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

PKC iota/lambda/zeta Antibody [Unconjugated] - Cross-reactive Summary

• Immunogen: E. coli-derived recombinant human PKC iota
  Ile455-Val596
  Accession # P41743
• Specificity: Detects human, mouse and rat PKC iota / lambda / zeta in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for PKC iota/lambda/zeta Antibody [Unconjugated] - Cross-reactive

• PKC iota/lambda/zeta

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PKC iota/lambda/zeta Antibody (AF4465)(1)

Publication using AF4465

• AC Engevik, ES Krystofiak, I Kaji, AR Meyer, VG Weis, A Goldstein, AW Coutts, T Melkamu, M Saqui-Salc, JR Goldenring Recruitment of polarity complexes and tight junction proteins to the site of apical bulk endocytosis Cellular and Molecular Gastroenterology and Hepatology, 2021-02-03;0(0):. 2021-02-03 [PMID: 33548596] (IHC, Mouse)

Review for PKC iota/lambda/zeta Antibody (AF4465) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer IP Mouse 07/31/2015

Summary

• Application: Immunoprecipitation
• Sample Tested: Hippocampus
• Species: Mouse

FAQs for PKC iota/lambda/zeta Antibody (AF4465). (Showing 1 - 1 of 1 FAQ).

1. Does this antibody work as an IP antibody to pull-down PKCiota/lambda and their phosphorylated form?
   • For product AF4465, this item has not yet been tested in an immunoprecipitation assay. This antibody is manufactured by our sister company R&D Systems and has only been tested so far in western blot and IHC per the R&D Systems site. The immunogen of the antibody is found between aa 455-596 of the human Protein kinase C iota type protein. This sequence encompasses one phosphorylation site (aa 564), and therefore has the potential to recognize the phosphorylated and non-phosphorylated forms of the protein; with exception of p Thr 564. This, of course, is hypothetical as the antibody has not yet been tested against the phosphorylated forms of the proteins.

Bioinformatics

• Uniprot:
  • P41743()