Thermo Scientific™ Pierce™ EDC, No-Weigh™ Format

Catalog No.	Quantity
PIA35391	10 x 1 mg
PI77149	10 mg
PI22980	5 g
PI22981	25 g

Catalog No. PIA35391 Supplier Thermo Scientific™ Supplier No. A35391

Description

Immobilize or conjugate proteins using 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (CAS 25952-53-8), a water-soluble, carboxyl-to-amine crosslinker.

Thermo Scientific™ Pierce EDC is a water-soluble carbodiimide crosslinker that activates carboxyl groups for spontaneous reaction with primary amines, enabling peptide immobilization and hapten-carrier protein conjugation.

Thermo Scientific No-Weigh products are specialty reagents provided in pre-aliquoted format. The pre-weighed packaging helps prevent loss of reagent reactivity and contamination over time by minimizing repetitive opening and closing of vial. The format enables use of fresh vial of reagent each time, eliminating hassle of weighing small amounts of reagents and reducing concerns over reagent stability.

• Reactive group: Carbodiimide
• Reaction target: Activates carboxyl groups to conjugate to amino groups (primary amines)
• Several conjugation strategies: React EDC alone with target groups or include NHS or Sulfo-NHS to increase reaction efficiency or to stabilize active intermediate for later reaction to amines
• Neutral linkage: Forms neutral amide bonds between carboxyls and amines
• Water-soluble Reagent: Add directly to reactions in aqueous, physiological buffers
• Soluble reaction byproducts: Easily removed by washing with water or dilute acid
• High-purity, crystalline reagent: Use to create high-quality, activated derivatives

Applications:

• Conjugate carboxyl and amino groups among peptides and proteins
• Couple haptens to immunogenic carrier proteins (e.g., attach a peptide to KLH)
• Immobilize peptide antigens to affinity-purify antibodies
• Create NHS-activated, amine-reactive labeling compounds
• Crosslink proteins to carboxyl coated beads or surfaces
• Activate nanoparticles with amine-reactive Sulfo-NHS esters

Specifications

• Chemical Reactivity: Amine-Carboxyl
• Cleavable: No
• Description: EDC
• Molecular Weight (g/mol): 191.7
• PEGylated: No
• Spacer Arm Length: 0.0 Å
• Content And Storage: Upon receipt store desiccated at -20°C.
• Cell Permeability: No
• Shipping Condition: Wet Ice
• Product Line: No-Weigh, Pierce
• Labeling Method: Chemical Labeling
• Crosslinker Type: Heterobifunctional
• Reactive Moiety: Carbodiimide
• Spacer: Short (<10 Å)
• Form: Powder
• Quantity: 10 x 1 mg
• Solubility: Water
• Format: Standard, Single-use, Premium-grade
• Water Soluble: Yes
• Product Type: Crosslinker

Frequently Asked Questions (FAQs)

Can NHS and EDC stock solutions be made for long-term storage before use?

No. EDC is very unstable in aqueous environments and must be dissolved immediately before use. NHS and Sulfo-NHS are relatively stable in solution but best results are obtained when they are dissolved immediately before use. Store these compounds desiccated at 4°C.

How are NHS and Sulfo-NHS used?

These compounds are used to modify a carboxyl group to form an amine-reactive ester. This is accomplished by mixing NHS with a carboxyl-containing molecule and the carbodiimide EDC (Cat. No. 22980, 22981, 77149, A35391). EDC causes a dehydration reaction between the carboxyl and the NHS hydroxyl group, giving rise to an NHS-ester-activated molecule. The activated molecule can then be reacted spontaneously with a primary amine-containing molecule. Although the carboxyl-molecules can be made to react directly with amines using EDC, the reaction is much more efficient with NHS because a stable intermediate is created. In fact, molecules that are activated as NHS esters can be dried and stored for later reaction to amine-containing targets.

What are NHS and Sulfo-NHS used for?

These compounds are used in conjunction with the crosslinker EDC (Cat. No. 22980, 22981, 77149, A35391) to synthesize amine-reactive labeling reagents, crosslinkers and conjugation compounds. Any compound containing a carboxylic acid (-COOH), such as a protein, or biotin or peptide, can be activated with NHS or Sulfo-NHS to form an NHS ester that will spontaneously react to form covalent amide linkages with proteins and other molecules that contain primary amines (-NH2).

What is the difference between the Thermo Scientific CarboLink (now Glycolink) and CarboxyLink Resins?

The CarboxyLink Resin is for immobilizing carboxyl-containing biomolecules after EDC activation. CarboLink Resin is hydrazide-activated and immobilizes glycoproteins. Carbolink has been replaced with glycolink immobilization kits and resins. Glycolink can also be used to immobilize steroids or other molecules containing ketones. Both resins are capable of coupling ligands via carboxylic acids using EDC (Cat. No. 22980). Both immobilization chemistries are available on UltraLink Resin as UltraLink Hydrazide (glycoproteins) and UltraLink DADPA (carboxyl) Resins.

How do CarboxyLink and DADPA UltraLink Resins immobilize molecules?

These resins are derivatized with diaminodipropylamine (DADPA). DADPA has a terminal primary amine that enables covalent immobilization of peptides or other carboxyl-containing (-COOH) molecules. When incubated with the resin and the carbodiimide crosslinker EDC (included in the CarboxyLink Immobilization Kit), carboxyl-containing molecules become permanently attached to the resin by stable amide bonds. The immobilized molecules can then be used in affinity purification procedures. CarboxyLink/DADPA UltraLink Coupling Resins can also be used to immobilize other kinds of molecules using alternative amine-reactive crosslinking chemistries.

Can you provide the shelf-life for EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride)?

EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.

Is your EDAC compound the same as your EDC compound?

Yes, both acronyms are used for the same compound. The chemical name for EDC is 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and the chemical name for EDAC is N-3-(3-dimethylaminopropyl)-N-ethyl-carbodiimide. Both of these compounds are structurally equivalent.

What are the general characteristics of N-hydroxysuccinimide esters (NHS Esters)?

NHS esters are reactive groups formed by EDC-activation of carboxylate molecules. NHS ester-activated crosslinkers and labeling compounds react with primary amines in slightly alkaline conditions (pH 7.2-8.5) to yield stable amide bonds. The reaction releases N-hydroxysuccinimide (MW 115), which can be removed easily by dialysis or desalting. Primary amine buffers such as Tris or TBS are not compatible because they compete for reaction; however, in some procedures, it is useful to add Tris or glycine buffer at the end of a conjugation procedure to quench (stop) the reaction.
Sulfo-NHS esters are identical to NHS esters except that they contain a sulfonate (–SO3) group on the N-hydroxysuccinimide ring. This charged group has no effect on the reaction chemistry, but it does tend to increase the water-solubility of crosslinkers containing them. In addition, the charged group prevents sulfo-NHS crosslinkers from permeating cell membranes, enabling them to be used for cell surface crosslinking methods.

What are the general characteristics of carbodiimides?

EDC and other carbodiimides are zero-length crosslinkers; they cause direct conjugation of carboxylates (-COOH) to primary amines (-NH2) without becoming part of the final crosslink (amide bond) between target molecules. EDC crosslinking reactions must be performed in conditions devoid of extraneous carboxyls and amines. Because peptides and proteins contain multiple carboxyls and amines, direct EDC-mediated crosslinking usually causes random polymerization of polypeptides. Nevertheless, this reaction chemistry is used widely in immobilization procedures (e.g., attaching proteins to a carboxylated surface) and in immunogen preparation (e.g., attaching a small peptide to a large carrier protein).

What is the difference between GlycoLink and CarboxyLink Gels?

GlycoLink gel is hydrazide-activated crosslinked beaded agarose, and it is useful for coupling glycoproteins via aldehydes formed from their sugars by sodium meta-periodate oxidation. Reaction of aldehydes with hydrazide- activated resin is catalyzed by aniline resulting in >90% coupling in 4 hours or less. CarboxyLink gel on the other hand is crosslinked beaded agarose activated with diaminodipropylamine (DADPA) and is useful for immobilizing carboxyl- containing biomolecules after EDC activation. Actually, both resins can be used with EDC to couple ligands via carboxylic acids.

Note: Both immobilization chemistries are available on UltraLink Support as UltraLink Hydrazide and UltraLink DADPA respectively.

What is the difference between SulfoLink and CarboxyLink Coupling Resin supports?

SulfoLink Coupling Resin is iodoacetyl-activated agarose that reacts with reduced thiols to form a stable thioether linkage. CarboxyLink Coupling Resin is an amine-derivatized agarose that utilizes the carbodiimide crosslinker, EDC to conjugate the carbon of carboxylic acid to the nitrogen of the immobilized primary amine (hydrazide) to form a permanent amide bond.

What are the benefits of using a CarboxyLink support?

CarboxyLink can be used to immobilize any protein or peptide via the C-terminus or aspartic or glutamic acid residues. Diaminodiproylamine (DADPA) is immobilized onto an agarose support. The carbodiimide crosslinker, EDC facilitates the formation of an amide bond between the carbon of the carboxylic acid and the nitrogen on the terminal amine of DADPA. This support is compatible with 50% organic solvent to improve immobilization of hydrophobic peptides.

For Research Use Only. Not for use in diagnostic procedures.