IMMPACT:Initiative for Cell Mortality Programme Assessment.

G Protein Pathways, Part B: G Proteins and their Regulators

Cross-Linking Concanavalin A to SPR Chips

Concanavalin A is a tetrameric protein with four identical 28 kDa subunits. 29 The minimal structure exhibiting high affinity binding of carbohydrates is the dimeric form. The dimer form of ConA is an active stable structure, and the tetramer – dimer transition can be promoted by low pH or succinylation of the protein.30 For reasons of economy we use unmodified concanavalin A Clia (Genprice Inc.). Because the dissociation of dimer from tetramer leads to a continual decline in the SPR baseline, our coupling procedures first promote the tetramer – dimer dissociation prior to linkage. To accomplish this, ConA is dissolved and stored at 4 ° in 100 mM sodium acetate, pH 4.8, prior to coupling. This also provides for optimal concentration of the ConA dimer by ionic attraction to the unactivated carboxyl groups. For limiting the ConA coupling the pH can be adjusted to 5.4 and still promote the dimer form.

However, extensive washing of the surface to allow complete tetramer dissociation is then necessary. Linkage of the ConA is accomplished by standard NHS / EDC amine coupling16 as described for streptavidin above, but with a modification in the running solution (Solution C, 50 mM MOPS, pH 7.5, 3 mM MgSO4, 150 mM NaCl with 10 μM CaCl2 and 10 μM MnCl2.

First, the surface is activated by injection of a mixture of 0.2 M EDC with 50 mM NHS in water at a flow rate of 5 μl / min, exposure time of 5–7 min. This is followed by injection of ConA lectin at 0.1–0.3 mg / ml in 100 mM sodium acetate, pH 4.8, with an exposure time of 5–7 min. Activated carboxyls are then quenched by reaction with 1 M ethanolamine in water with an exposure time of 4 min. These procedures lead to the immobilization predominantly of dimeric ConA.

The immobilized ConA is quite stable, so that rhodopsin coupling does not have to proceed immediately. However, the coupling reactions are sufficiently reproducible and rapid that we find no advantage to prior preparation of ConA-modified surfaces. For experiments defining the independent binding of Gα or Gβγ described below, surfaces modified with 7000–12,000 RU of Biomag plus ConA have been utilized.

Under these conditions, the presence of immobilized tetrameric ConA is a significant factor. When high concentrations of ConA and / or aggressive NHS / EDC activation conditions are used, we routinely wash the ConA surface for 30–60 min with Solution C at a flow rate of 5 μl / min to allow for complete tetramer – dimer dissociation, prior to rhodopsin immobilization.

Concanavalin A (ConA)
05-0106-1000	Medicago	1 g	EUR 238.8

Concanavalin A (ConA)
05-0106-10000	Medicago	10 g	Ask for price

Concanavalin A (ConA)
05-0106-100000	Medicago	100 g	Ask for price

Concanavalin A (ConA)
05-0106-250	Medicago	250 mg	EUR 105.6

BioMagPlus Concanavalin A
BP531-10	Bangs Laboratories	10 ml	EUR 1399.73
Description: BioMagPlus Concanavalin A is magnetic particles with affinity binding proteins may be coated with the primary antibody or biotinylated molecule

BioMagPlus Concanavalin A
BP531-3	Bangs Laboratories	3 ml	EUR 621.95
Description: BioMagPlus Concanavalin A is magnetic particles with affinity binding proteins may be coated with the primary antibody or biotinylated molecule

Concanavalin A ELISA Kit
20-abx052181	Abbexa	EUR 7726.80 EUR 4118.40 EUR 961.20	10 × 96 tests 5 × 96 tests 96 tests

Concanavalin A (Con A) Antibody
20-abx101152	Abbexa	EUR 326.40 EUR 159.60 EUR 811.20 EUR 410.40 EUR 292.80	100 ug 10 ug 1 mg 200 ug 50 ug

Concanavalin A (ConA) CLIA Kit
20-abx190017	Abbexa	EUR 9174.00 EUR 4888.80 EUR 1130.40	10 × 96 tests 5 × 96 tests 96 tests

Concanavalin A, CF350 conjugate
29015	Biotium	5x1mg	EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF633 conjugate
29018	Biotium	5x1mg	EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF680 conjugate
29020	Biotium	5x1mg	EUR 288
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF770 conjugate
29058	Biotium	5x1mg	EUR 288
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF750 conjugate
29080	Biotium	5x1mg	EUR 288
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF594 conjugate
29017	Biotium	5x1mg	EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Rat Concanavalin A ELISA kit
E02C0795-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Rat Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rat Concanavalin A ELISA kit
E02C0795-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Rat Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Rat Concanavalin A ELISA kit
E02C0795-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Rat Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Concanavalin A ELISA kit
E08C0795-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Canine Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Concanavalin A ELISA kit
E08C0795-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Canine Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Dog Concanavalin A ELISA kit
E08C0795-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Canine Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Concanavalin A ELISA kit
E07C0795-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Porcine Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Concanavalin A ELISA kit
E07C0795-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Porcine Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Pig Concanavalin A ELISA kit
E07C0795-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Porcine Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Concanavalin A (ConA) ELISA Kit
20-abx150329	Abbexa	EUR 7256.40 EUR 3873.60 EUR 904.80	10 × 96 tests 5 × 96 tests 96 tests

Concanavalin A, CF488A conjugate
29016	Biotium	5x1mg	EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF640R conjugate
29019	Biotium	5x1mg	EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF405M conjugate
29074	Biotium	5x1mg	EUR 262.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF405S conjugate
29075	Biotium	5x1mg	EUR 262.8
Description: Minimum order quantity: 1 unit of 5x1mg

Goat Concanavalin A ELISA kit
E06C0795-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Goat Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat Concanavalin A ELISA kit
E06C0795-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Goat Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat Concanavalin A ELISA kit
E06C0795-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Goat Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human Concanavalin A ELISA kit
E01C0795-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Human Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human Concanavalin A ELISA kit
E01C0795-48	BlueGene	1 plate of 48 wells	EUR 624
Description: A competitive ELISA for quantitative measurement of Human Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human Concanavalin A ELISA kit
E01C0795-96	BlueGene	1 plate of 96 wells	EUR 822
Description: A competitive ELISA for quantitative measurement of Human Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Mouse Concanavalin A ELISA kit
E03C0795-192T	BlueGene	192 tests	EUR 1524
Description: A competitive ELISA for quantitative measurement of Mouse Concanavalin A in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.