ALEXIS Biochemicals: PARP-1 (bovine) (automodified) Standard

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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-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-202-024

Revised 24-May-07

Calmodulin (bovine brain) (high purity)
SYNONYMS	CaM (bovine brain) (high purity) Phosphodiesterase 3':5'-cyclic Nucleotide Activator (bovine brain) (high purity)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Calmodulins

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-202-024-M001		1 mg	180.00 USD

Product Specification

CAS NUMBER:	73298-54-1
MERCK INDEX:	14: 1719
SOURCE/HOST:	Isolated from bovine brain.
PURITY:	≥95% (native reducing PAGE in presence of EGTA)
PURITY DETAIL:	Ion-exchange chromatography followed by gel filtration.
FORMULATION:	Lyophilized. Contains 2mM EDTA
RECONSTITUTION:	Reconstitute with distilled water.
SPECIFIC ACTIVITY:	~40'000U/mg protein. One unit is defined as the amount of enzyme that stimulates 0.016 activated units of phosphodiesterase 3’:5’-cyclic nucleotide (Prod. No. ALX-202-035) to 50% of the maximum activity of the enzyme when saturated with activator, in the presence of 0.01mM Ca²⁺ at 30°C, pH 7.5.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	-20°C
HANDLING:	Avoid freeze/thaw cycles.

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P02593: Calmodulin (human)
AfCS Signalling Gateway link A000452: Calmodulin I (mouse)

Further Categories Containing This Product:

Natural Proteins

ALX-200-005

Revised 12-Apr-07

Calmodulin (human brain) (high purity)
SYNONYMS	CaM (human brain) (high purity) Phosphodiesterase 3':5'-cyclic Nucleotide Activator (human brain) (high purity)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Calmodulins

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-200-005-C500		500 µg	135.00 USD

Product Specification

CAS NUMBER:	73298-54-1
MERCK INDEX:	14: 1719
SOURCE/HOST:	Isolated from human brain. Tested negative for infectious diseases.
PURITY:	≥95% (native reducing PAGE in presence of EGTA)
PURITY DETAIL:	Ion-exchange chromatography and gel filtration.
FORMULATION:	Lyophilized. Contains 2mM EDTA.
RECONSTITUTION:	Reconstitute with distilled water.
SPECIFIC ACTIVITY:	~40'000U/mg protein. One unit is defined as the amount of calmodulin that stimulates 0.016 activated units of phosphodiesterase 3':5'-cyclic nucleotide (Prod. No. ALX-202-035) to 50% of the maximum activity of the enzyme when saturated with activator, in the presence of 0.01mM Ca²⁺at 30°C, pH 7.5.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	-20°C

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P02593: Calmodulin (human)
AfCS Signalling Gateway link A000452: Calmodulin I (mouse)

Further Categories Containing This Product:

Natural Proteins

ALX-202-062

Revised 16-Jun-08

Calmodulin (pig)
SYNONYMS	CaM (pig) Phosphodiesterase 3’:5’-cyclic Nucleotide Activator (pig)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Calmodulins

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-202-062-M001		1 mg	75.00 USD
ALX-202-062B-C100	Biotin	100 µg	220.00 USD
ALX-202-062L-C100	Fluorescein	100 µg	220.00 USD
ALX-202-062RH-C100	Rhodamine	100 µg	220.00 USD

Product Specification

MW:	~17kDa (SDS-PAGE)
MERCK INDEX:	14: 1719
SOURCE/HOST:	Isolated from pig brain.
QUANTITY:	ALX-202-062B: biotin:protein = 0.5-1.0
PURITY:	≥98% (SDS-PAGE)
FORMULATION:	Lyophilized from 5% sucrose, pH 7.0-7.4, containing 10mM sodium phosphate.
RECONSTITUTION:	Reconstitute with distilled water to 1mg/ml.
SPECIFIC ACTIVITY:	≥35’000U/mg protein. One unit will stimulate 0.01 unit of 3’:5’-cyclic nucleotide phosphodiesterase in a 3ml reaction volume at pH 7.5 and 30°C, to 50% of the maximum activity of the enzyme when saturated with CaM in the presence of 100µM Ca²⁺.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	-20°C
HANDLING:	Protect from light. Avoid freeze/thaw cycles.

Product Specific Literature References

Trifluoperazine binding to porcine brain calmodulin and skeletal muscle troponin C: L. Massom, et al.; Biochemistry 29, 671 (1990) Abstract

Further Categories Containing This Product:

Natural Proteins

ALX-204-005

Revised 07-Aug-07

Calmodulin (wheat)
SYNONYMS	CaM (wheat) Phosphodiesterase 3':5'-cyclic Nucleotide Activator (wheat)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Calmodulins

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-204-005-C250		250 µg	70.00 USD
ALX-204-005-M001		1 mg	190.00 USD
ALX-204-005B-C050	Biotin	50 µg	290.00 USD
ALX-204-005L-C050	Fluorescein	50 µg	290.00 USD
ALX-204-005RH-C050	Rhodamine	50 µg	290.00 USD

Product Specification

MW:	~17kDa (SDS-PAGE)
MERCK INDEX:	14: 1719
SOURCE/HOST:	Isolated from wheat germ (Triticum aestivum).
PURITY:	≥98% (SDS-PAGE)
FORMULATION:	Lyophilized from 5% sucrose, pH 7.0, containing 10mM sodium phosphate.
RECONSTITUTION:	Reconstitute with 1ml distilled water.
SPECIFIC ACTIVITY:	≥35’000U/mg protein. One unit will stimulate 0.01 unit of 3’:5’-cyclic nucleotide phosphodiesterase in a 3ml reaction volume at pH 7.5 and 30°C, to 50% of the maximum activity of the enzyme when saturated with CaM in the presence of 100µM Ca²⁺.
APPLICATION:	Unconjugated CaM: Can be independently derivatized at both N- and C-terminal regions within the molecule for structural, proteomic biomarker discovery and protein-protein interaction studies. CaM-Biotin, -Fluorescein & -Rhodamine: Protein array applications for the detection of calmodulin binding proteins (CBPs), Western blot and others.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	-20°C
HANDLING:	Protect from light. Avoid freeze/thaw cycles.

Product Description

Wheat germ calmodulin sequence is unique in that it contains cysteine (Cys²⁶) and only a single tyrosine (Tyr¹³⁸), in contrast to mammalian CaMs, which have two tyrosine residues and lack cysteine. This allows wheat germ CaM to be independently derivatized at both N- and C-terminal regions. Therefore it is ideal for applications requiring both calmodulin activation and residue specific tagging. Wheat germ CaM also has no tryptophan resulting in a distinctive absorbance spectrum when compared to mammalian CaM sequences. The conjugated formats are labelled at Cys²⁶.

Product Specific Literature References

Site-specific derivatives of wheat germ calmodulin. Interactions with troponin and sarcoplasmic reticulum: G.M. Strasburg, et al.; J. Biol. Chem. 263, 542 (1988) Abstract; Full Text

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P04464: Calmodulin (wheat)

General Literature References

A rapid and sensitive method for detection and quantification of calcineurin and calmodulin-binding proteins using biotinylated calmodulin: M.L. Billingsley, et al.; PNAS 82, 7585 (1985) Abstract; Full Text

Further Categories Containing This Product:

Natural Proteins

ALX-212-014

Revised 30-Jan-07

Calmodulin (wheat & pig) (Immobilized High Loading)
SYNONYMS	CaM (wheat & pig) (Immobilized High Loading) Phosphodiesterase 3’:5’-cyclic Nucleotide Activator (wheat & pig) (Immobilized High Loading)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Calmodulins

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-212-014-R200		200 µl	290.00 USD

Product Specification

MERCK INDEX:	14: 1719
SOURCE/HOST:	Isolated from wheat germ (Triticum aestivum) and pig brain. Immobilized via SH group (Cys²⁶) (wheat) and amino group (pig) to epoxy activated resin (non-agarose).
CONCENTRATION:	Calmodulin content: 1ml of resin bound with 5-5.5mg of wheat germ calmodulin.
FORMULATION:	50% slurry in 3M NaCl, 20mM sodium phosphate buffer, pH 7.4 and 1mM EGTA.
APPLICATION:	Useful for pull down experiments and proteomic applications requiring a high capacity precipitation reagent. Dense resin can be easily separated from extracts by centrifugation using batch or spin column procedures. Resin can be reused but proteolysis from crude samples will reduce binding capacity on repeat cycles. Loading conditions: 100µM Ca²⁺, with protease inhibitors and appropriate buffers. Elution conditions: Buffers with 1mM EDTA or EGTA.
SHIPPING:	SHIPPED ON BLUE ICE
LONG TERM STORAGE:	+4°C

Product Description

Wheat germ calmodulin sequence is unique in that it contains cysteine (Cys²⁶) and only a single tyrosine (Tyr¹³⁸), in contrast to mammalian CaMs, which have two tyrosine residues and lack cysteine. Immobilized via SH group (Cys²⁶) (wheat) and amino group (pig) to epoxy activated resin (non-agarose). Thus allowing multiple orientations of bound calmodulin for efficient capture of calmodulin binding proteins.

Product Specific Literature References

Site-specific derivatives of wheat germ calmodulin. Interactions with troponin and sarcoplasmic reticulum: G.M. Strasburg, et al.; J. Biol. Chem. 263, 542 (1988) Abstract; Full Text

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P04464: Calmodulin (wheat)

Further Categories Containing This Product:

Natural Proteins

ALX-212-013

Revised 30-Jan-07

Calmodulin (wheat) (Immobilized High Loading)
SYNONYMS	CaM (wheat) (Immobilized High Loading) Phosphodiesterase 3’:5’-cyclic Nucleotide Activator (wheat) (Immobilized High Loading)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Calmodulins

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-212-013-R200		200 µl	290.00 USD

Product Specification

MERCK INDEX:	14: 1719
SOURCE/HOST:	Isolated from wheat germ (Triticum aestivum). Coupled primarily through SH group (Cys²⁶) to epoxy activated resin (non-agarose).
CONCENTRATION:	Calmodulin content: 1ml of resin bound with 5-5.5mg of wheat germ calmodulin.
FORMULATION:	50% slurry in 3M NaCl, 20mM sodium phosphate buffer, pH 7.4