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14-497 Sigma-AldrichPKC η Protein, active, 10 µg

Active, N-terminal His6-tagged, recombinant amino acids 2-end, of human PKC eta. For use in Kinase Assays.

PKC η Protein, active, 10 µg MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

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Description
Catalogue Number	14-497
Brand Family	Upstate
Trade Name	Upstate
Description	PKC η Protein, active, 10 µg
Overview	N-terminal His6-tagged, recombinant amino acids 2-end, of human PKC eta. PKC Lipid activator (Catalog # 20-133) is required for use.

References

Product Information
Quality Level	MQ100

Applications
Application	Active, N-terminal His6-tagged, recombinant amino acids 2-end, of human PKC eta. For use in Kinase Assays.
Key Applications	Kinase Assay

Biological Information
Source	Human PKC eta expressed in Sf21 cells.
Specific Activity	For Specific Activity data, refer to the Certificate of Analysis for individual lots of this enzyme.
Entrez Gene Number	NM_006255.3
Entrez Gene Summary	Protein kinase C (PKC) is a family of serine- and threonine-specific protein kinases that can be activated by calcium and the second messenger diacylglycerol. PKC family members phosphorylate a wide variety of protein targets and are known to be involved in diverse cellular signaling pathways. PKC family members also serve as major receptors for phorbol esters, a class of tumor promoters. Each member of the PKC family has a specific expression profile and is believed to play a distinct role in cells. The protein encoded by this gene is one of the PKC family members. It is a calcium-independent and phospholipids-dependent protein kinase. It is predominantly expressed in epithelial tissues and has been shown to reside specifically in the cell nucleus. This protein kinase can regulate keratinocyte differentiation by activating the MAP kinase MAPK13 (p38delta)-activated protein kinase cascade that targets CCAAT/enhancer-binding protein alpha (CEBPA). It is also found to mediate the transcription activation of the transglutaminase 1 (TGM1) gene.
Gene Symbol	PRKCH PRKCL PKCL MGC26269 nPKC-eta MGC5363 PKC-L
Protein Target	PKCη
Purification Method	Ni²⁺/NTA-agarose
Target Sub-Family	AGC
UniProt Number	P24723
UniProt Summary	FUNCTION: SwissProt: P24723 # PKC is activated by diacylglycerol which in turn phosphorylates a range of cellular proteins. PKC also serves as the receptor for phorbol esters, a class of tumor promoters. SIZE: 682 amino acids; 77563 Da TISSUE SPECIFICITY: Most abundant in lung, less in heart and skin. DOMAIN: SwissProt: P24723 The C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor and the C2 domain is a non-calcium binding domain. DISEASE: SwissProt: P24723 # Defects in PRKCH may be a cause of susceptibility to ischemic stroke [MIM:601367]; also known as cerebrovascular accident or cerebral infarction. A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors. SIMILARITY: Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily. & Contains 1 AGC-kinase C-terminal domain. & Contains 1 C2 domain. & Contains 2 phorbol-ester/DAG-type zinc fingers. & Contains 1 protein kinase domain.
Molecular Weight	78.6kDa

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	Routinely evaluated by phosphorylation of PKC Substrate Peptide
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	6 months at -70°C

Packaging Information
Material Size	10 µg
Material Package	Also available in 250 µg size --call for pricing and availability and reference catalog number 14-497M when ordering the 250 µg size.

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Katalogové číslo	GTIN
14-497	04053252674617

Documentation

PKC η Protein, active, 10 µg MSDS

Title
Safety Data Sheet (SDS)

PKC η Protein, active, 10 µg Certificates of Analysis

Title	Lot Number
PKC eta, active - 25609U-B	25609U-B

FAQ

Question	Answer
Why is it important to shake the reaction mixture during the incubations?	It is important to shake the reaction mixture during incubations so that the substrate and the enzyme get good interaction. If you do not have access to a shaking incubator, use a water bath for the incubations, removing the tubes regularly to vortex.
What type of buffer is best for lysis when attempting to preserve a protein?	The best buffer is Buffer A. It is composed of 50mM Tris/HCl, pH 7.5, 1mM EGTA, 1mM EDTA, 1mM Sodium Orthovanadate (activated), 10mM Na B-glycerolphosphate, 50mM NaF, 5mM Na pyrophosphate, 1% w/v Triton X-100, 0.1% w/v 2-mercaptoethanol, 1ug/ml each of aprotinin, leupeptin and pepstatin, 1mM PMSF and 1mM microcystin (this is optional). QS with water. You may store Buffer A in aliquots at -20°C for 6 months.
What is the Molar extinction coefficient for Adenosine 5'-[g32P]triphosphate ([g32P]ATP) used in the kinase assay?	The Molar extinction coefficient is 15400.
What is the best way to store and thaw enzymes?	In order to maintain activity of enzymes, we recommend a slow thaw and aliquot storage. To do this, remove the full vial from the freezer and place under cold running tap water. Once thawed, place the vial on ice, gently mix, than make aliquots. Using liquid nitrogen or an ethanol/dry ice bath, flash freeze the aliquots. Store aliquots at the recommended temperature.
When using p81 paper (phosphocellulose squares), what is the purpose of the acetone wash?	The purpose of washing the p81 squares in acetone is mostly to help them dry faster, but it also can help to eliminate low specificity binding.
Some of my enzyme preparations require Brij-35, what is its purpose?	The role of Brij-35 is to function as a mild detergent, which prevents aggregation in active kinases.

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Kategorie

Life Science Research > Proteins and Enzymes > Purified Kinases

Life Science Research > Drug Discovery and Development > Kinase & Phosphatase Screening > Purified Kinases

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