供货周期: | 一周 |
品牌: | |
规格: | 1μg/3μg/100μg |
货号: | PKA-204 |
CAS号: |
PKA-RII alpha
cAMP-Dependent Protein Kinase A regulatory subunit-II A Recombinant
Catalog No. PKA-204
Synonyms:
cAMP-Dependent Protein Kinase A regulatory subunit-II A, PKA-RII alpha.
Introduction:
cAMP-dependent PKA is a ubiquitous serine/theonine protein kinase present in a variety of tissues (e.g. brain, skeletal muscle, heart). The intracellular cAMP level regulates cellular responses by altering the interaction between the catatytic C and regulatory R subunits of PKA. The inactive tetrameric PKA holoenzyme R2C2 is activated when cAMP binds to R2, which dissociates the tetramer to R2 cAMP 4 and two active catalytic subunits. Free Catalytic subunits of PKA can phosphorylate a wide variety of intracellular target proteins. In response to hormone- induced high cAMP levels, PKA phosphorylates glycogen synthetase (inhibition of the enzyme activity) and phosphorylase kinase to block glycogen synthesis. Different isoforms of catalytic and regulatory subunits suggest specific functions.
Description:
The recombinant PKA regulatory subunit II-a is a dimeric 90 kDa protein.
Protein Kinase A is purified by proprietary chromatographic techniques.
Source:
Escherichia Coli.
Physical Appearance:
Sterile Filtered clear solution.
Formulation:
PKA regulatory subunit-II alpha is supplied in 50% glycerol.
Unit Definition:
One unit is defined as the amount of recombinant PKA catalytic subunit alpha, required to incorporate 1nmol of phosphate into the specific substrate peptide kemptide (LRRASlG) in one minute at 30°C.
Stability:
PKA should be stored at 4°C if entire vial will be used within 2-4 weeks. For long term storage it is recommended to store at -20°C.
Avoid multiple freeze-thaw cycles.
Purity:
Greater than 95% as determined by SDS-PAGE.
Biological Activity:
PKA regulatory subunit alpha specifically inhibits PKA catalytic subunit (Ki about 0.7nM). The binding is not dependent on the presence of ATPMg, however, the regulatory subunit will be phosphorylated at the binding site by PKA C alpha upon binding. Activity can be restored by adding the second messenger cAMP (Kact about 100nM). Subcellular localisation is regulated via interaction of PKA RII with so-called A-kinase anchoring proteins (AKAPs).
Assay Conditions:
Roskoski-Assay:
Protein kinase activity can be measured using a modified radioactive assay according to Roskoski et al.: The assay will be performed in a mixture containing 50mM MOPS (pH7.0), 10mM MgCI2, 0.25 mg/ml bovine serum albumin, 100 IJM Kemptide (peptide substrate), 100 IJM unlabeled ATP mixed with [y_32p] ATP (500-1000 cpm/pmol) and Ca subunit in a final volume of 50 IJI. Reaction is started by addition of the Ca subunit and can be stopped after a 5 minute incubation at 30°C by spotting the reaction mix onto Whatman P-81 filters and soaking the filters four times in 75mM phosphoric acid (10 ml per sample) for at least 5 minutes. After four washing steps rinse filters with ethanol, dry and count.
Roskoski, R., Jr. (1983) Methods Enzymol. 99, 3-6
For the detection of phosphorylation in substrate proteins the phosphotransferase reaction can alternatively be stopped by taking aliquots of the mixture and adding SDS sample buffer. The phosphorylation status of the substrate proteins can subsequently be analysed using SDS PAGE and autoradiography.
Zimmermann, B. (1999) Journal of Biological Chemistry.274, 9, 5370-78
Usage:
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
相关产品
重组人肌球蛋白轻链2(Recombinant Human MYL2)
重组大鼠抵抗素(Rat Resistin)
大鼠酸性磷酸酶(ACP)ELISA Kit
大鼠胆囊收缩素/缩胆囊素八肽(CCK-8)ELISA Kit
人瘦素(LEP)ELISA Kit
重组人半乳糖凝集素16/LGALS16 Human
重组人半乳糖凝集素8/Human LGALS8 His
重组人半乳糖凝集素3/Human LGALS3 His
重组人卵泡抑素/Human FST His
重组人β防御素118/DEFB118 Human
重组人心肌营养素1/CTF1 Human
特级新生牛血清(无支原体)
超级新生牛血清(无支原体)
优级胎牛血清(无支原体)
特级新生牛血清(无噬菌体低内毒素)
详细地址
关注
拨打电话
留言咨询