Phospho-PKC (pan) (gamma Thr514) (PF4) rabbit mAb

Catalog #: 2451
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200 uL (0.5 mg/mL)(2451) $475.00
20 uL (0.5 mg/mL)(2451S) $175.00
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Note

20 μL is enough antibody for at least 20 Western blots.

Product Overview

Description

Protein Kinase (PKC) is a 12 member family of serine/threonine kinases termed conventional or classical  (a b, g), novel (d,e,and q), atypical (zl) and PKN and PKC-related (PKN1, PKN2 and PKN3) playing significant role in several signaling processes involved in physiological and pathological setting (1). PKC activation translates into gene expression modulation, cell division, migration, proliferation, differentiation, and cell survival and apoptosis (2).  PKC members are classified based on their distinct cofactor requirements and the extent of homology between their regulatory elements (3,4). PKCalpha, beta I, beta II, and gamma constitute the conventional PKC isoforms, characterized by the presence of two cysteine-rich zinc finger domains, C1a and C1b (5,6) which bind to diacylglycerol (DAG) and phosphatidylserine (PS) (7). In addition, PKC contains a C2 domain, responsible for binding anionic phospholipids like phosphatidylinositol bisphosphate (PIP2) in a Ca2+-dependent manner (8,9). The atypical PKC, PKCz andl share ATP-binding domain and the catalytic domain with PKC. They contain a single C1 domain that lacks residues necessary for binding DAG (10).

 Association of PKCg to the membrane enables a conformation that permits phosphoinositide-dependent protein kinase1 (PDK-1) (11) to bind and phosphorylate a site in the activation loop, Thr514. (12,13). Thr514 phosphorylation leads to a conformation change enabling phosphorylation of at two carboxyl-terminal sites namely, the turn motif and hydrophobic motif, as a result of which the fully phosphorylated conventional PKC is released from the membrane, and positioned in the cytoplasm as an inactive form (14,15). Binding of Ca2+ induces low-affinity interaction with the membrane, whereas the membrane imbedded cofactor DAG to PKC results in high-affinity interaction of PKC with the membrane (16). 

Product Details

Product Details
Specification Description
Applications Flow Cytometry, WB
Clone PKCgT514-PF4
Format Unconjugated
Validated Reactivity Human, Mouse, Rat
Cross Reactivity Predicted to work with mouse, rat and other homologues.
Detection Anti-Rabbit IgG
Clonality Monoclonal
Immunogen A synthetic phospho-peptide corresponding to residues surrounding Thr514 of human phospho PKCγ
Formulation 1X PBS, 0.02% NaN3, 50% Glycerol, 0.1% BSA
Isotype Rabbit IgGk
Preparation Protein A+G
Recommended Usage 1µg/mL – 0.001µg/mL. It is recommended that the reagent be titrated for optimal performance for each application. See product image legends for additional information.
Storage -20ºC
Pseudonyms Protein kinase C gamma type, PRKCG, PKCG
Uniprot ID P17252 P05771 P05771-2 Q05655 Q02156 P24723 P05129 Q04759
References
1. Mugami S, et al., 2018, Mol. Cell. Endocrinol. 463:97-105.
2. Isakov N, 2018, Semin. Cancer Biol. 48:36-52.
3. Mellor H, and Parker PJ, 1998, Biochem. J. 332:281-92.
4. Newton AC, 1995, J. Biol. Chem. 270:28495-8.
5. Ho C, et al., 2001, Biochem. 40:10334-41.
6. Hurley JH, et al., 1996, Protein Sci. 6:477-80.
7. Colon-Gonzalez F, and Kazanietz MG, 2006, Biochim. Biophys. Acta 1761 :827-37.
8. Naleski EA, and Falke JJ, 1996, Protien Sci, 5 :2375-90.
9. Orr JW, and Newton AC, 1992, Biochem 31:4667-4673.
10. Antal CE and Newton AC, 2014, Biochem Soc. Trans. 42:1477-1483.
11. Sonnenberg ED, et al., 2001 J Biol Chem, 276:45289-97.
12. Dephoure N, et al., 2008, Proc. Natl. Acad Sci. U.S.A. 105:10762-67.
13. Barnett ME, et al., 2007, Cell Signal. 19:1820-9.
14. Newton AC, 2003, Biochem. J. 370 :361-71.
15. Parekh DB, et al., 2000, EMBO J. 10:496-503.
16. Edwards AS, et al., 1999, J Biol. Chem., 274:6461-8.

Images

Cell Culture Products
Flow cytometric analysis of HT1080 cells treated with staurosporine (red) or untreated (green) using Phospho-PKCg (Thr514) (PF4) Rabbit mAb. at 0.05 ug/mL, PKCgT514-PF4 #2451, or concentration-matched Rabbit (G9) mAb IgG Isotype Control #2141 for cells treated with staurosporine (black) or untreated (blue). Flow cytometric analysis of HT1080 cells, treated with staurosporine and stained with the secondary antibody only as negative control (blue) or treated with staurosporine (red) or untreated (green) using Phospho-PKC (pan) (gamma Thr514) antibody PKCgT514-PF4 at 0.05 µg/mL. Cat. #2451.
Cell Culture Products
Flow cytometric analysis of Jurkat cells, treated with K252a and stained with the secondary antibody only negative control (blue) or treated with K252a (red) or untreated (green) using Phospho-PKC (pan) (gamma Thr514) antibody PKCgT514-PF4 at 0.1 µg/mL. Cat. #2451.
Cell Culture Products
Peptide blocking flow cytometric analysis of HT1080 cells secondary antibody only negative control (light blue) or untreated (red) or treated with staurosporine (green) or untreated and blocked with phospho-peptide (black) or treated with staurosporine and blocked with phospho peptide (gold) or untreated and blocked with non-phospho peptide (dark blue) or treated with staurosporine and blocked with non-phospho peptide (purple) using Phospho-PKCg (Thr514) antibody PKCGT514-PF4 at 0.05 µg/mL. Cat. #2451.
Cell Culture Products
Western blot analysis of NIH3T3 cell extract untreated or treated with TPA using Phospho-PKC (pan) (gamma Thr514) antibody PKCgT514-PF4 at 0.05 ug/mL. Cat. #2451.
Cell Culture Products
Western blot analysis of C6 cell extract untreated or treated with staurosporine using Phospho-PKC (pan) gamma (Thr514) antibody PKCgT514-PF4 at 0.1 ug/mL. Cat. #2451.

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