immobilized tetrameric Concanavalin A

concanavalin a cona

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 (Sigma). 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.

concanavalin a
concanavalin a

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 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 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)

05-0106-100000 100 g Ask for price

Concanavalin A (ConA)

05-0106-250 250 mg
EUR 105.6

Concanavalin A (ConA)

E16FXP010 1 mg
EUR 86.67
Description: Available in various conjugation types.

BioMagPlus Concanavalin A

BP531-10 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 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
  • EUR 7726.80
  • EUR 4118.40
  • EUR 961.20
  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Concanavalin A (Con A) Antibody

20-abx101152
  • 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 Lectin (Con A) - Cy3

21761096-1 -
EUR 148.5

Concanavalin A Lectin (Con A) - Cy5

21761160-1 -
EUR 142.41

Concanavalin A Lectin (Con A) - Pure

22070010-1 -
EUR 111

Concanavalin A Lectin (Con A) - Pure

22070010-3 -
EUR 72.06

Concanavalin A Lectin (Con A) - Pure

22070010-4 -
EUR 104.25

Concanavalin A (ConA) CLIA Kit

20-abx190017
  • EUR 9174.00
  • EUR 4888.80
  • EUR 1130.40
  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Concanavalin A (ConA) CLIA Kit

EKU08954-48T 48T
EUR 647.92

Concanavalin A (ConA) CLIA Kit

EKU08954-5x96T 5x96T
EUR 4396.6

Concanavalin A (ConA) CLIA Kit

EKU08954-96T 96T
EUR 925.6

Concanavalin A (ConA) CLIA Kit

EKN50065-48T 48T
EUR 403.06

Concanavalin A (ConA) CLIA Kit

EKN50065-5x96T 5x96T
EUR 2735.05

Concanavalin A (ConA) CLIA Kit

EKN50065-96T 96T
EUR 575.8

Concanavalin A, CF350 conjugate

29015 5x1mg
EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF633 conjugate

29018 5x1mg
EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF680 conjugate

29020 5x1mg
EUR 288
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF770 conjugate

29058 5x1mg
EUR 288
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF750 conjugate

29080 5x1mg
EUR 288
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF594 conjugate

29017 5x1mg
EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Rat Concanavalin A ELISA kit

E02C0795-192T 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 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 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 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 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 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 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 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 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, CF488A conjugate

29016 5x1mg
EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF640R conjugate

29019 5x1mg
EUR 250.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF405M conjugate

29074 5x1mg
EUR 262.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A, CF405S conjugate

29075 5x1mg
EUR 262.8
Description: Minimum order quantity: 1 unit of 5x1mg

Concanavalin A (ConA) ELISA Kit

20-abx150329
  • EUR 7256.40
  • EUR 3873.60
  • EUR 904.80
  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Concanavalin A (ConA) ELISA Kit

EKN44546-48T 48T
EUR 294

Concanavalin A (ConA) ELISA Kit

EKN44546-5x96T 5x96T
EUR 1995

Concanavalin A (ConA) ELISA Kit

EKN44546-96T 96T
EUR 420

Concanavalin A (ConA) ELISA Kit

EKU03435-48T 48T
EUR 431.69

Concanavalin A (ConA) ELISA Kit

EKU03435-5x96T 5x96T
EUR 2929.33

Concanavalin A (ConA) ELISA Kit

EKU03435-96T 96T
EUR 616.7

ConA(Concanavalin A) ELISA Kit

ELK1340-48T 48T Ask for price
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to ConA. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to ConA. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain ConA, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ConA in the samples is then determined by comparing the OD of the samples to the standard curve.

ConA(Concanavalin A) ELISA Kit

ELK1340-96T 96T Ask for price
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to ConA. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to ConA. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain ConA, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ConA in the samples is then determined by comparing the OD of the samples to the standard curve.