ALEXIS Biochemicals: cGMP-dependent Protein Kinase Iβ (human) (recombinant)

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ALX-202-027

Revised 09-Aug-06

cAMP-dependent Protein Kinase (bovine heart) (catalytic subunit)
SYNONYMS	Protein Kinase A (bovine heart) (catalytic subunit) PKA (bovine heart) (catalytic subunit)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PKA

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-202-027-U500		500 U	250.00 USD

Product Specification

MW:	~40kDa.
EC:	2.7.1.37
SOURCE/HOST:	Isolated from bovine heart.
CONCENTRATION:	20µg/ml.
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 20mM MES, pH 6.5, containing 0.1mM EDTA, 30mM β-mercaptoethanol, 100mM sodium chloride and 50% ethylene glycol.
SPECIFIC ACTIVITY:	~1000 Units/µg using histone as the substrate. One unit is defined as the amount of enzyme that transfers 1pmol phosphate to histone per min. at 30°C.
APPLICATION:	For studying PKA phosphorylation in vitro.
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C
HANDLING:	Do not freeze/thaw.

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link: cAMP-dependent Protein Kinase (bovine) α catalytic subunit (P00517), β-1 catalytic subunit (P05131), β-2 catalytic subunit (P24256)
AfCS Signalling Gateway link: cAMP-dependent Protein Kinase (mouse) α catalytic subunit (A001914), β catalytic subunit (A001815), γ catalytic subunit (A001916)

Further Categories Containing This Product:

Natural Proteins • Enzymes

ALX-201-081

Revised 30-Jun-06

Arginase l (human) (recombinant)
PRODUCT LINE	Nitric Oxide Pathway
PRODUCT CATEGORY	Arginine & Arginases / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-081-C020		20 µg	240.00 USD

Product Specification

EC:	3.5.3.1
SOURCE/HOST:	Produced in E. coli. Full length human arginase I.
CONCENTRATION:
PURITY:	≥90% (SDS-PAGE)
FORMULATION:	Liquid. In 10mM TRIS-HCl, pH 7.5, containing 1mM β-mercaptoethanol, 1mM MnCl₂ and 50% glycerol.
SPECIFIC ACTIVITY:	~2.0U/µg protein. One unit is defined as the amount of enzyme that converts 1µmol of L-arginine to L-ornithine and urea per min. at 37°C, pH 9.5 (R.T. Schminke, et al.; J. Biol. Chem. 238, 1012 (1962)).
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C

Product Specific Literature References

Determination of arginase activity in macrophages: a micromethod: I.M. Corraliza, et al.; J. Immunol. Methods 174, 231 (1994) Abstract
Implications of the S-shaped domain in the quaternary structure of human arginase: A. Mora, et al.; Biochim. Biophys. Acta 1476, 181 (2000) Abstract

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P05089: Arginase I (human)

Further Categories Containing This Product:

Enzymes • Recombinant Proteins / Fusion Proteins

ALX-208-002

Revised 03-Nov-08

C3 Exoenzyme
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	ADP Ribosylation Factor [ARF] / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-208-002-C010		10 µg	160.00 USD
ALX-208-002-C050		50 µg	390.00 USD

Product Specification

SOURCE/HOST:	Isolated from Clostridium botulinum.
CONCENTRATION:	1mg/ml after reconstitution.
PURITY:	≥95% (SDS-PAGE)
APPEARANCE:	Lyophilized from 5mM HEPES, pH 8.0.
RECONSTITUTION:	Reconstitute with 10µl/50µl distilled water.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	-20°C
HANDLING:	After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles.

Product Description

Exhibits ADP-ribosyltransferase activity. Not neurotoxic. Not cell permeable. The substrate for C3 exoenzyme has been identified as Rho, a GTP-binding Ras-related protein. Rac proteins are approximately 100-fold less active as substrates. The ADP-ribosylation site of exoenzyme C3 on rho A has been identified as Asn⁴¹.

Product Specific Literature References

ADP-ribosylation of platelet actin by botulinum C2 toxin: K. Aktories, et al.; Eur. J. Biochem. 161, 155 (1986) Abstract
K. Aktories, et al.; TIPS 8, 158 (1987)
Clostridium botulinum type C produces a novel ADP-ribosyltransferase distinct from botulinum C2 toxin: K. Aktories, et al.; FEBS Lett. 212, 109 (1987) Abstract
Botulinum C2 toxin ADP-ribosylates actin and disorganizes the microfilament network in intact cells: K.H. Reuner, et al.; Eur. J. Cell Biol. 43, 134 (1987) Abstract
Botulinum ADP-ribosyltransferase C3. Purification of the enzyme and characterization of the ADP-ribosylation reaction in platelet membranes: K. Aktories, et al.; Eur. J. Biochem. 172, 445 (1988) Abstract
Functional modification of a 21-kilodalton G protein when ADP- ribosylated by exoenzyme C3 of Clostridium botulinum: E.J. Rubin, et al.; Mol. Cell Biol. 8, 418 (1988) Abstract
The rho gene product expressed in E. coli is a substrate of botulinum ADP-ribosyltransferase C3: K. Aktories, et al.; BBRC 158, 209 (1989) Abstract
K. Aktories and A. Hall; TIPS 10, 415 (1989) Abstract
The mammalian G protein rhoC is ADP-ribosylated by Clostridium botulinum exoenzyme C3 and affects actin microfilaments in Vero cells: P. Chardin, et al.; EMBO J. 8, 1087 (1989) Abstract
Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase: A. Sekine, et al.; J. Biol. Chem. 264, 8602 (1989) Abstract; Full Text
Microinjection of recombinant p21rho induces rapid changes in cell morphology: H.F. Paterson, et al.; J. Cell. Biol. 111, 1001 (1990) Abstract
Characterization of the C3 gene of Clostridium botulinum types C and D and its expression in Escherichia coli: A. Popoff, et al.; Infect. Immun. 59, 3673 (1991) Abstract
The ras protein family: evolutionary tree and role of conserved amino acids: A. Valencia, et al.; Biochemistry 30, 4637 (1991) Abstract
ADP-ribosylation of the ras-related, GTP-binding protein RhoA inhibits lymphocyte-mediated cytotoxicity: P. Lang, et al.; J. Biol. Chem. 267, 11677 (1992) Abstract; Full Text
A rho gene product in human blood platelets. II. Effects of the ADP- ribosylation by botulinum C3 ADP-ribosyltransferase on platelet aggregation: N. Morii, et al.; J. Biol. Chem. 267, 20921 (1992) Abstract; Full Text
The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors: A.J. Ridley, et al.; Cell 70, 389 (1992) Abstract
Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility: K. Takaishi, et al.; Mol. Cell. Biol. 13, 72 (1993) Abstract
Effect of botulinum C3 exoenzyme on cell growth and cytoskeleton organization in transformed human epidermal cells in culture: a possible role for rho protein in epidermal cells: M. Yamamoto, et al.; J. Dermatol. Sci. 8, 103 (1994) Abstract

General Information

All proteins encoded by the superfamily of ras genes are structurally related, bind GTP and exhibit GTPase activity which is regulated by divalent cations. In human cells, three slight variations of the Rho protein (Rho A, B and C, respectively) have been described.

BACKGROUND/TECHNICAL INFORMATION

APD-ribosylation is performed as described by Chardin et al. Target G proteins or subcellular fractions corresponding to 50-70µg protein are incubated for 1 hour at 37°C with 20-30ng of C3 in a 10mM HEPES buffer, pH 8.0 containing, 15mM isonicotinic acid hydrazide, 15mM thymidine, 1mM MgCl₂, 1mM ATP, 2µM [³²P]NAD. The reaction is stopped by addition of 0.06M TRIS-HCl, pH 7.0, 15% (v/v) glycerol, 5% β-mercaptoethanol, 2.3% SDS and 0.001% bromophenol blue. The mixture may then be subjected to SDS-PAGE.

Further Categories Containing This Product:

Enzymes • Natural Proteins

ALX-202-026

Revised 30-Aug-07

Ca2+/Calmodulin-dependent Protein Kinase II (rat brain)
SYNONYMS	CaM Kinase II (rat brain) CAMK2 (rat brain)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	CAM Kinases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-202-026-C001		1 µg	295.00 USD

Product Specification

MW:	~65kDa. Composed of 60kDa and 50kDa subunits in ratios of 1:3 to 1:4.
EC:	2.7.1.123
SOURCE/HOST:	Isolated from rat brain. Requires activation.
CONCENTRATION:	20µg/ml
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 25mM TRIS/HCl, pH 7.5, 0.5mM EDTA, 10mM β-mercaptoethanol, 10µM AEBSF, 50% glycerol and 0.1mg/ml leupeptin.
RECONSTITUTION:	Reconstitute with sterile distilled water.
SPECIFIC ACTIVITY:	~1µmol/mg/min at 30°C using synapsin I as the substrate.
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C
HANDLING:	Avoid freeze/thaw cycles.

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link: Ca²⁺/Calmodulin-dependent Protein Kinase II (rat) α chain (P11275), β chain (P08413), γ chain (P11730), δ chain (P15791)
AfCS Signalling Gateway link: Ca²⁺/Calmodulin-dependent Protein Kinase II (mouse) α chain (A000456), β chain (A000457), γ chain (A000459), δ chain (A000458)

Further Categories Containing This Product:

Natural Proteins • Enzymes

ALX-202-034

Revised 13-Dec-04

Calcineurin (bovine brain)
SYNONYMS	Calmodulin Binding Protein (bovine brain) Protein Phosphatase 2B (bovine brain)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PP2B [Calcineurin] / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-202-034-U100		100 U	590.00 USD

Product Specification

CAS NUMBER:	9025-75-6
EC:	3.1.3.16
MERCK INDEX:	14: 1640
SOURCE/HOST:	Isolated from bovine brain.
FORMULATION:	Lyophilized powder containing ~1% protein (Lowry) and 0.5% EGTA, buffer salts and stabilizers.
SPECIFIC ACTIVITY:	~2'500-5'000U/mg protein. One unit causes a 50% inhibition of the activated phosphodiesterase activity when assayed with 2 units of activator and 0.1mM Ca²⁺ in an enzyme coupled system at 30°C, pH 7.5.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	-20°C

Further Categories Containing This Product:

Enzymes • Natural Proteins • Nitric Oxide Pathway Other Products

ALX-201-224

Revised 22-Feb-06

cAMP-dependent Protein Kinase (Catalytic Subunit Cα) (mouse) (recombinant)
SYNONYMS	Protein Kinase A (Catalytic Subunit Cα) (mouse) (recombinant) PKA (Catalytic Subunit Cα) (mouse) (recombinant)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PKA

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-224-C025		25 µg	305.00 USD
ALX-201-224-C050		50 µg	474.00 USD
ALX-201-224-C100		100 µg	812.00 USD

Product Specification

MW:	~41kDa
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 25mM potassium phosphate, pH 6.5, containing 5mM β-mercaptoethanol, 5mM EDTA, 150mM NaCl and 50% glycerol.
SPECIFIC ACTIVITY:	1.5x10⁷U/mg. One unit is defined as the amount of enzyme required to incorporate 1pmol phosphate per min into the specific substrate kemptide (Prod. No. ALX-160-006) at 30°C.
APPLICATION:	- In vitro enzymological studies of neural and hormonal signal transduction - Phosphorylation of target proteins in vivo (via microinjection) including ion channels, transcriptional activator proteins and regulatory enzymes of glycogen metabolism
SHIPPING:	SHIPPED ON DRY ICE
SHORT TERM STORAGE:	-20°C
LONG TERM STORAGE:	-80°C
USE/STABILITY:	Stable for 6 months after receipt when stored at -20°C. Stable for 2 weeks when stored at +4°C.
HANDLING:	Avoid freeze/thaw cycles.

Further Categories Containing This Product:

Enzymes • Recombinant Proteins / Fusion Proteins

ALX-201-229

Revised 15-May-07

cAMP-dependent Protein Kinase (Holoenzyme Type IIα, Cα2 (mouse)/RIIα2 (human)) (recombinant)
SYNONYMS	Protein Kinase A (Holoenzyme Type IIα, Cα₂ (mouse)/RIIα₂ (human)) (recombinant) PKA (Holoenzyme Type IIα, Cα₂ (mouse)/RIIα₂ (human)) (recombinant)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PKA

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-229-C020		20 µg	474.00 USD
ALX-201-229-C050		50 µg	812.00 USD
ALX-201-229-C100		100 µg	1'489.00 USD

Product Specification

MW:	~172kDa
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:	~1mg/ml
FORMULATION:	Liquid. In 20mM MOPS, pH 7.0, containing 150mM sodium chloride and 50% glycerol.
SPECIFIC ACTIVITY:	1.5x10⁷U/mg (for the Cα catalytic subunit of the activated holoenzyme). One unit is defined as the amount of enzyme required to incorporate 1pmol phosphate per min into the specific substrate kemptide (Prod. No. ALX-160-006) at 30°C.
APPLICATION:	Suitable for analyzing PKA type II agonists (cAMP analogs) or antagonists.
SHIPPING:	SHIPPED ON DRY ICE
SHORT TERM STORAGE:	-20°C
LONG TERM STORAGE:	-80°C
USE/STABILITY:	Stable for at least 6 months after receipt when stored at -20°C.
HANDLING:	Avoid freeze/thaw cycles.

Product Description

Inactive holoenzyme consisting of one dimeric human regulatory subunit type IIα and two monomeric mouse catalytic subunits. Activate by addition of the second messenger cAMP (K_a~100nM) (Prod. No. ALX-480-011) releasing two monomeric catalytic subunits.

Product Specific Literature References

[1] Assays of protein kinase: R. Roskoski, Jr.; Meth. Enzymol. 99, 3 (1983) Abstract
[2] PrKX is a novel catalytic subunit of the cAMP-dependent protein kinase regulated by the regulatory subunit type I: B. Zimmermann, et al.; J. Biol. Chem. 274, 5370 (1999) Abstract; Full Text

General Information

BACKGROUND/TECHNICAL INFORMATION

Roskoski-Assay

Protein kinase activity may be measured by using a modified radioactive assay [1].
The assay is performed in a mixture containing 50mM MOPS, pH 7.0, 10mM MgCI₂, 0.25mg/ml BSA, 100µM kemptide (peptide substrate), 100µM unlabeled ATP mixed with [γ-³²p] ATP (500-1000cpm/pmol) in a final volume of 50µI. Reaction is started by addition of the Cα subunit from activated PKA holoenzyme, type IIα and can be stopped after a 5 min. incubation at 30°C by spotting the reaction mix onto Whatman^® P-81 filters and soaking the filters four times in 75mM phosphoric acid (10ml per sample) for at least 5 min. After four washing steps rinse filters with ethanol, dry and count.
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 [2].

Further Categories Containing This Product:

Enzymes • Recombinant Proteins / Fusion Proteins

ALX-201-225

Revised 26-Jul-05

cAMP-dependent Protein Kinase (Regulatory Subunit RIα) (bovine) (recombinant)
SYNONYMS	Protein Kinase A (Regulatory Subunit RIα) (bovine) (recombinant) PKA (Regulatory Subunit RIα) (bovine) (recombinant)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PKA

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-225-C025		25 µg	474.00 USD
ALX-201-225-C050		50 µg	812.00 USD
ALX-201-225-C100		100 µg	1'489.00 USD

Product Specification

MW:	~43kDa/subunit; homodimer
SOURCE/HOST:	Produced in E. coli.
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 10mM potassium phosphate, pH 6.8, containing 150mM NaCl, 14mM 2-mercaptoethanol, 1mM EDTA and 50% glycerol.
BIOLOGICAL ACTIVITY:	Reversibly inhibits the catalytic subunit Cα of cAMP-dependent protein kinase (PKA) (Prod. No. ALX-201-224). The inhibition of the catalytic subunit can be reversed by the addition of the second messenger cAMP (>1µM) (Prod. No. ALX-480-011).
SHIPPING:	SHIPPED ON DRY ICE
SHORT TERM STORAGE:	-20°C
LONG TERM STORAGE:	-80°C
USE/STABILITY:	Keep the glycerol stock solution of the enzyme permanently at -20°C while pipetting to avoid loss of inhibitory activity.
HANDLING:	Avoid freeze/thaw cycles.

Further Categories Containing This Product:

Enzymes • Recombinant Proteins / Fusion Proteins

ALX-201-226

Revised 18-Jan-06

cAMP-dependent Protein Kinase (Regulatory Subunit RIIα) (human) (recombinant)
SYNONYMS	Protein Kinase A (Regulatory Subunit RIIα) (human) (recombinant) PKA (Regulatory Subunit RIIα) (human) (recombinant)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PKA

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-226-C025		25 µg	474.00 USD
ALX-201-226-C050		50 µg	812.00 USD
ALX-201-226-C100		100 µg	1'489.00 USD

Product Specification

MW:	45kDa/subunit; homodimer
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 10mM potassium phosphate, pH 6.8, containing 150mM NaCl, 14mM β-mercaptoethanol, 1mM EDTA and 50% glycerol.
BIOLOGICAL ACTIVITY:	Reversibly inhibits the catalytic subunit Cα of cAMP-dependent protein kinase (PKA) (Prod. No. ALX-201-224). The inhibition of the catalytic subunit can be reversed by the addition of the second messenger cAMP (>1µM) (Prod. No. ALX-480-011).
APPLICATION:	Suitable for the analysis of interactions with so-called A-kinase anchoring proteins (AKAPs).
SHIPPING:	SHIPPED ON DRY ICE
SHORT TERM STORAGE:	-20°C
LONG TERM STORAGE:	-80°C
USE/STABILITY:	Keep the glycerol stock solution of the enzyme permanently at -20°C while pipetting to avoid loss of inhibitory activity.
HANDLING:	Avoid freeze/thaw cycles.

Further Categories Containing This Product:

Enzymes • Recombinant Proteins / Fusion Proteins

ALX-201-227

Revised 14-Nov-07

cAMP-dependent Protein Kinase (Regulatory Subunit RIIβ) (rat) (recombinant)
SYNONYMS	Protein Kinase A (Regulatory Subunit RIIβ) (rat) (recombinant) PKA (Regulatory Subunit RIIβ) (rat) (recombinant)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	PKA

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-227-C025		25 µg	474.00 USD
ALX-201-227-C050		50 µg	812.00 USD
ALX-201-227-C100		100 µg	1'489.00 USD

Product Specification

MW:	45kDa/subunit; homodimer
SOURCE/HOST:	Produced in E. coli.
CONCENTRATION:
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 10mM potassium phosphate, pH 6.8, containing 150mM NaCl, 14mM 2-mercaptoethanol, 1mM EDTA and 50% glycerol.
BIOLOGICAL ACTIVITY:	Reversibly inhibits the catalytic subunit Cα of cAMP-dependent protein kinase (PKA) (Prod. No. ALX-201-224). The in