Biochemistry of Proteins

DETERGENTS: SOLUBILIZATION OF BIOMOLECULES

See also: Dialysis - Protein dosage (detergent compatibility)

 

Uptima offers a range of highly purified detergent for requirements of :

  • Protein solubilization

  • Immunological techniques

  • Biochemistry of lipids and proteins

Uptima purified detergents are recommended each time that biomolecule activity (receptors, enzymes…) and analysis (electrophoresis, bioassay…) may be affected by the quality of detergents. This occurs with no proteomic grade detergents, notably especially for polyoxyethylenes and carbohydrates derivatives.

  1. Detergents, general information

  2. Complete list of detergents

  3. Oxidant free detergents

  4. Carbohydrate derived detergents

 

Detergents, general information

Detergents are water soluble molecules classified according to their hydrophilic/hydrophobic character and ionic groups. This effectively drives the pattern of protein/detergents interactions. Notably their hydrophobic tail associates to form micelles, or aggregates, or interacts with other molecules (lipids, proteins).

Proteins are usually extracted by lysis of the cells and tissues in presence of detergents (SDS, Triton® X100, X114, CHAPS, DOC, NP40, OctylThioGlucosides) that desorganize the lipidic bilayer of membranes, and solubilized proteins. Non solubilized material is harvested by centrifugation or other means.

In solution, detergents help keeping molecules in solution, by dissociating aggregates, increasing solubility, and unfolding proteins.

In electrophoresis notably, proteins are classically treated with SDS to denature the native tertiary and quaternary structures, allowing the separation of proteins according to their molecular weights.

Uptima offers detergents of the highest quality to meet the proteomic grade for extraction, purification, storage, and electrophoresis analysis.

Physical properties of useful detergents

Cat.#

Detergent

Type

PM

Aggregation Number 

Micelle PM

CMC (mM)

HLB (mM)

Cloud pt (°C)

Common Applications

UP09187

Brij-35

non-ionic

1225

40

49 k

0,05-(0.09)-0.1

16.9

>100

SMP, Chrom&Electr.

UP31473

Brij-56

non-ionic

a

a

a

a

a

a

a

UP31474

Brij-58

non-ionic

1120

70

82 k

0,077

15.7

>100

a

-

Digitonin

non-ionic

1229.3

5-6

a

a

a

a

a

UP01838

Guanidine

-

-

-

-

-

-

-

-

UP78548

Hecameg

non-ionic

335.4

a

a

19.5

a

a

SMP (receptors)

24367

Nonidet P-40

non-ionic

602

149

90 k

0.5-(0.29)-0.3

13.1

80

SPM ; Chrom&Electr.

UP60207

n-nonyl-b-D-glucopyranoside

non-ionic

306.4

a

a

6.5

a

a

a

UP26370

n-octyl-b-D-glucopyranoside

non-ionic

292.4

84

7.9 k

20-25

a

>100

SMP, Anal., Immunol..

UP60208

n-octyl-b-D-thioglucopyranoside

non-ionic

308.4

a

a

9

a

>100

SMP (mild), Enz.

UP42613

n-decyl-b-D-maltopyranoside

non-ionic

482.6

a

a

1.8

a

a

a

UP34675

n-dodecyl-b-D-maltopyranoside

non-ionic

510.6

98

a

0.1-(0.17)-0.6

a

a

SMP, Anal., Enz., Chrom&Electr.

UP55205

n-undecyl-b-D-maltopyranoside

non-ionic

496.6

a

a

0.59

a

a

a

UP52112

Triton X-100

non-ionic

Av.646

100-(140)-155

90 k

0.2-(0,23)-0.35

13.5

64

SMP, all purposes

UP15852

Triton X-114

non-ionic

Av.536

a

a

0.17

12.4

23

SMP, Enz.

UP15874

Tween 20

non-ionic

Av.1227

a

a

0.059

16.7

95

Immunol. Solub.Anal

UP15878

Tween 80

non-ionic

Av.1310

60

76 k

0.012

15

a

Immunol.Stabil.IgM

UP33351

CHAPS

zwitterionic

614.9

4-14

6.1k

6-(8)-10

-

>100

SMP, Anal., Enz., Chrom&Electr.

UP35639

CHAPSO

zwitterionic

630.9

10

6149

8

a

>100

SMP, Anal., Enz., Chrom&Electr.

UP11708

DOC

anionic

414.6

5

2 k

1.5

-

-

-

UP64910

SDS

-

-

-

-

-

-

-

-

UP89682

SDS

anionic

288.5

62

18-24k

7-10

40

>100

Electroph., SMP

UP03190

Urea

a

60.06

a

a

a

a

a

Electroph.

Notes: SMP = Sample Preparation, Enz.=enzymatic studies, Anal.=Analytical techniques, Chrom&Electr=Chromatography and Electrophoresis analysis.

Oxidant free detergents

A lot of polyoxyethylenic detergents are available in a variety of structures and under trade names, like famous Brij®, Tween® and Triton®. Most are industrial products containing a mixture of related structures. The composition often varies with batches. When the manufacturing is of bad quality, several contaminants impair biological activity of biomolecules like enzymes, labile tertiary structures, receptors… Furthermore, polyoxyethylenic detergents tend to alter, mostly by oxidation, and this aging is increased by light and temperature. Oxidation generates peroxides, carboxyls and free radicals that may reach up to 0.2%.

Even at far lower concentration, these oxidizing agents may be very critical in your application: Such contaminants are often responsible for inactivation, denaturation, fragmentation of the desired biomolecule (Jaeger 1994), especially during the extraction from cell lysates. Sulfhydryls are readily oxidized by peroxides that induce aggregations of receptors (Chang 1974) or transition of conformation state (O’Brien 1975). Contaminants may also interfere in biochemical analysis (i.e. for estrogen (Lever 1977), or even protein with Coomassie (Stuzenbberg 1992).

Do you want reliables results? Choose Uptima oxydant free detergents. They are purified by chromatography, and contain less than 5-20 µM of peroxides equivalents. They are supplied as convenient 10% solution amps.

Brij-35

UP091870

5x10ml

Brij-56

UP314736

5x10ml

Oxidant Free, 10% solution

UP091871

10x10ml

Oxidant Free, 10% solution

UP314737

10x10ml

23 Lauryl ether ; C12E23 ;

CAS: 9002-92-0

Peroxide content (HPLC) : < 20 %

Main Applications:

sample preparation, chromatography

10 Cethyl ether

Peroxide content (HPLC) : < 20 %

Brij-58

UP314740

5x10ml

Oxidant Free, 10% solution

UP314741

10x10ml

20 Cethyl ether

Peroxide content (HPLC) : < 20 %

Triton X-100

UP521121

5x10ml

Triton X-305

UP708534

5x10ml

Oxidant Free, 10% solution

UP521122

10x10ml

Oxidant Free, 10% solution

UP708535

10x10ml

Octyl phenoxy poly ethoxy phenol

CAS: 9002-93-1

Peroxide content (HPLC) : < 20 %

Main Applications:

Protein extraction

Peroxide content (HPLC) : < 20 %

Triton X-114

UP158528

5x10ml

Triton X-405

UP158536

5x10ml

Oxidant Free, 10% solution

UP158529

10x10ml

Oxidant Free, 10% solution

UP158537

10x10ml

CAS: 9036-19-5

Peroxide content (HPLC) : < 20 %

Main Applications:

Protein extraction

Peroxide content (HPLC) : < 20 %

Tween 20

UP158740

5 x10ml

Tween 80

UP158780

5x10ml

Oxidant Free, 10% solution

UP158741

10x10ml

Oxidant Free, 10% solution

UP158781

10x10ml

Polyoxyethylene sorbitan ; C12-sorbitan-E20

Peroxide content (HPLC) : < 20 %

Main Applications:

Immunoassays, surfactant

Polyoxyethylene sorbitan ; C18:1-sorbitan-E20

Peroxide content (HPLC) : < 20 %

Main Applications:

IgM stabilization, vaccin ingredient, emulsifier

CHAPS, UltraPure

UP333514

5g

CHAPSO, UltraPure

UP356392

1g

UP33351A

25g

UP356393

5g
UP333515 100g

(3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate)

Zwitterionic detergent (electronically neutral)

Protects the native state of proteins

Easily removed by dialysis

  • Purity (HPLC) > 99%

  • Conductivity (0.5 M) < 50 µS

 

Security Sheet

(3-[(3-Cholamidopropyl)dimethylammonio]-2-hydroxy

-1-propanesulfonate)

Zwitterionic detergent (electronically neutral)

Protects the native state of proteins

Easily removed by dialysis

  • Purity (HPLC) > 99%

  • pH (solution 1%) 5-8

  • Conductivity (0.5 M) <100 µS

SDS, UltraPure

UP896826

500ml at 20%

DOC, UltraPure

UP11708D

10g

Lauryl sulfate

UP896827

2 x 500ml at 20%

UP11708E

50g

Sodium Dodecyl Sulfate

Anionic detergent 

Excellent batch to batch reproducibility

Suitable for biotechnology, proteomics and genomics

  • Purity (HPLC) > 99%

  • Content in C12 : > 99%

  • Nuclease, RNase and protease free

  • OD260 and OD280 (3% solution in water) < 0.1

  • Chlorides < 0.1%

  • Copper, Lead < 5ppm

Deoxycholate

CAS: 302-95-4

MW: 414.57

  • Purity: > 99%

  • Heavy metals: <0.005%

Devoided of contaminants that may affect :

  • The renaturation of proteins (C16)

  • The detection sensitivity (UV absorbing subst.)

  • Separations and enzymatic activities (Heavy metals and chlorides)

Guanidine HCI

UP018380

500g

Hecameg

UP785480

5g

  • Purity (HPLC) > 99%

  • Chlorides, Sulfates : < 0.01%

  • OD 260 : < 0.03

  • OD 280 : < 0.15

  • As Pb Fe content : < 0.0005%

  • DNase, RNase, Proteases : non detected

(6-O-(N-heptylcarbamoyl)-methyl-b-D-glucopyranoside)

  • Purity (HPLC) > 99%

  • Non ionic

  • Useful for membrane proteins purification

Technical Sheet

Urea - UltraPure

UP031903

500g

UP031904 1kg

  • Purity (HPLC) > 99,5%

  • DNase, RNase, Proteases : non detected

  • OD 280 (8M in water) : < 0.15

  • Heavy metals content : < 0.001%

  • Solubility