Biotinylation Reagents

Biotin-Streptavidin is among the strongest interactions found in nature. Biotinylation reagents are widely used to enable the isolation, separation, concentration and further downstream processing or analysis of biomolecules. Appropriate biotinylation of the target molecule is essential to ensure high binding efficiency to streptavidin or Immobilized Streptavidin. ProteoChem manufactures an assortment of biotinylation reagents that target various functional groups like primary amines, sulfhydryls (thiols), carboxyls, and carbohydrates.

 

ProteoChem’s biotinylation reagents are offered as water soluble (Sulfo-NHS ester containing) or insoluble varieties. ProteoChem also offers biotins with diverse spacer arm lengths which can be used to reduce steric hindrances associated with streptavidin binding, thus allowing for more efficient binding of biotinylated proteins and peptides.

 

 

Biotinylation Reagent Selection Guide

 

 

 

1.  NHS-Biotin

Succinimidyl Biotin (NHS-Biotin) is a primary amine reactive biotinylation reagent. The NHS-Biotin molecule is cell membrane permeable and can be used to label intracellular proteins.

 

•  Molecular Weight: 341.38

•  Spacer Length: 13.5 Å

•  CAS Number: 35013-72-0

•  Alternative Names: Biotin-OSU; Biotin N-hydroxysuccinimide ester; (+)-Biotin N-succinimidyl ester

 

☞   NHS-Biotin Protocol & Product Information Sheet

 

 

2.  NHS-LC-Biotin

NHS-LC-Biotin, Succinimidyl 6-(biotinamido)hexanoate, is an amino-reactive biotinylation reagent with a long spacer arm. The 6-atom spacer arm of NHS-LC-Biotin provides distance between the biotin and the target ligand which helps alleviate steric hindrance. NHS-LC-Biotin is used to biotinylate proteins, peptides, or amino acids by reacting with primary amines as found in the N-terminus and Lysine side chains. The NHS-LC-Biotin molecule is cell membrane permeable and can be used to label intracellular proteins; whereas Sulfo-NHS-LC-Biotin is designed to biotinylate extracellular proteins. The resulting biotinylated protein can then be detected when used in conjunction with streptavidin and Immobilized Streptavidin.

 

•  Molecular Weight: 454.54

•  Spacer Length: 22.4 Å

•  CAS Number: 72040-63-2

•  Alternative Name: (+)-Biotinamidocaproate N-hydroxysuccinimidyl ester 

 

☞   NHS-LC-Biotin Protocol & Product Information Sheet

 

 

3.  NHS-SS-Biotin

Succinimidyl-2-(biotinamido)-ethyl-1,3'-dithiopropionate (NHS-SS-Biotin) is an amine-reactive biotinylation reagent that is cleavable when the disulfide (-S-S-) bond is reduced. NHS-SS-Biotin is cell membrane permeable and has a long chain separating the biotin group from the amino-reactive NHS-ester (N-Hydroxysuccinimide ester).

It is possible to cleave the biotin group from the crosslinked target protein under reducing conditions, such as in the presence of beta-mercaptoethanol (BME), DTT or TCEP. This technique is often used to purify an antibody or other protein from a biotin specific affinity resin like Immobilized Streptavidin without exposing your protein of interest to denaturing conditions. 

 

 

•  Molecular Weight: 504.65

•  Spacer Length: 24.3 Å

•  CAS Number: 142439-92-7

•  Alternate Names: Biotin disulfide N-Hydroxysuccinimide ester; NHS SS Biotin 

 

☞   NHS-SS-Biotin Protocol & Product Information Sheet

 

 

4.  Sulfo-NHS-Biotin

Sulfo-NHS-Biotin (Sulfosuccinimidyl biotin) is a water soluble biotinylation reagent and is packaged in 1 gram sizes for your large volume needs. Sulfo-NHS-Biotin is amine reactive and has a spacer arm length of 13.5 Å. Sulfo-NHS-Biotin is ideal for biotinylating antibody or protein that can then be used for protein detection or immobilization. Sulfo-NHS-Biotin is particularly useful for targeting cell surface proteins.

 

•  Molecular Weight: 443.4

•  Spacer Length: 13.5 Å

•  CAS Number: 119616-38-5

•  Alternative Names: Biotin 3-sulfo-N-hydroxysuccinimide ester sodium salt; Sulfosuccinimidobiotin 

 

☞   Sulfo-NHS-Biotin Product Information Sheet & General Protocol

 

 

 

5.  Sulfo-NHS-LC-Biotin

Sulfosuccinimidyl 6-(biotinamido) Hexanoate (Sulfo-NHS-LC-Biotin) is a long chain water soluble biotinylation reagent that is reactive toward primary amine groups and is packaged in 1 gram sizes for your large volume needs. Similar to Sulfo-NHS-Biotin (b2100-100mg and b2100-1gm), Sulfo-NHS-LC-Biotin is designed to target cell surface proteins. The sodium sulfonate groups lend water solubility to Sulfo-NHS-LC-Biotin and prevent it from crossing the lipid bi-layer of the cell membrane. 

 

 

•  Molecular Weight: 556.6

•  Spacer Length: 22.4 Å

•  CAS Number: 127062-22-0

•  Alternative Name: Biotinamidohexanoic acid 3-sulfo-N-hydroxysuccinimide ester sodium salt 

 

☞   Sulfo-NHS-LC-Biotin Product Information Sheet & General Protocol

 

 

 

6.  Sulfo-NHS-SS-Biotin

Sulfosuccinimidyl-2-(biotinamido)-ethyl-1,3'-dithiopropionate (Sulfo-NHS-SS-Biotin) is a cleavable, water-soluble, amino-reactive biotinylation reagent. This molecule is cell membrane impermeable due to its sodium sulfonate group. Sulfo-NHS-SS-Biotin has a long chain spacer arm separating the biotin group from the amino-reactive sulfo-NHS ester moity. It is possible to cleave the biotin group from the crosslinked target protein under reducing conditions, such as in the presence of BME, DTT or TCEP.

 

 

•  Molecular Weight: 606.69

•  Spacer Length: 24.3Å

•  CAS Number: 325143-98-4

•  Alternative Name: Biotin disulfide N-Hydroxysulfosuccinimide ester 

☞   Sulfo-NHS-SS-Biotin General Protocol & Product Information Sheet

 

 

7.  Biotin-HPDP

N-[6-(Biotinamido)hexyl]-3’-(2’-pyridyldithio)-propionamide (Biotin-HPDP) is a sulfhydryl reactive biotinylation reagent with a long chain, cleavable spacer arm. Biotin-HPDP biotinylation occurs between pH 6-9 through its pyridyldithiol group reacting with sulfhydryls to form a stable disulfide bond. The 29.2 angstrom spacer arm prevents steric hindrance and may also be cleaved at the disulfide bond using TCEP-HCl, DTT or ß-Mercaptoethanol. Biotin-HPDP must be dissolved in an organic solvent, such as DMSO or DMF, then added to an aqueous biotinylation reaction. The resulting biotinylated protein can then be detected when used in conjunction with streptavidin or Immobilized Streptavidin. 

 

 

•  Spacer Length: 29.2 Å

•  Molecular Weight: 539.78

•  CAS Number: 129179-83-5

•  Alternative Names: Biotin HPDP; Pyridyldithiol-biotin; Pyridyldisulfide-biotin 

 

 

 

8.  Iodoacetyl-LC-Biotin

Iodoacetyl-LC-Biotin (N-Iodoacetyl-N'-Biotinylhexylenediamine or N-Biotinyl-N'-(iodoacetyl)-1,6-hexanediamine) reacts with sulfhydryl groups to form stable thioether linkages. Iodoacetyl-LC-Biotin has an extended spacer arm that reduces steric hindrance. Iodoacetyl-LC-Biotin must be dissolved in an organic solvent, such as DMSO or DMF, then added to an aqueous biotinylation reaction. Iodoacetyl-LC-Biotin coupling reactions are most efficient when reacted at at pH 7.5 - 8.5 in reduced light. The resulting biotinylated protein can then be detected when used in conjunction with streptavidin or Immobilized Streptavidin.

 

 

•  Spacer Length: 27.1 Å

•  Molecular Weight: 510.43

•  CAS Number: 93285-75-7

•  Alternative Names: N-Biotinyl-N'-(iodoacetyl)-1,6-hexanediamine; N-iodoacetyl-N-biotinylhexylenediamine (IAB); Iaa-Biotin 

 

 

 

9.  Biotin Hydrazide

Biotin Hydrazide (Biotin-Hz) is a carbohydrate reactive biotinylation reagent. It is a great choice for labeling mouse monoclonal antibodies, unpaired cytosine residues in nucleic acids, and glycoprotiens.

•  Molecular Weight: 258.34

•  Spacer Length: 15.7 Å

•  CAS Number: 66640-86-6 

 

☞   Biotin Hydrazide Protocol & Product Information Sheet

 

 

 

10.  D-Biotin

D-Biotin, often called Vitamin H, D-(+)-Biotin, or simply Biotin, is water soluble. D-Biotin is often used in conjunction with Streptavidin or Immobilized Streptavidin.

 

•  Molecular Weight: 244.31

•  CAS Number: 58-85-5

•  Molecular Formula: C10H16N2O3S

•  MDL: MFCD00005541

•  EINECS: 200-399-3

•  Alternative Names: D-(+)-Biotin; Vitamin H; 5-(2-Oxo-hexahydro-thieno[3,4-d]imidazol-6-yl)-pentanoic acid; Bios II; Coenzyme R; Vitamin B7; Biotin

Cat. No	Product	Quantity
b2100-100mg	Sulfo-NHS-Biotin 100 mg	100 mg
b2100-1gm	Sulfo-NHS-Biotin 1 gram	1 gram
b2102-100mg	NHS-LC-Biotin 100 mg	100 mg
b2102-1gm	NHS-LC-Biotin 1 gram	1 gram
b2104-100mg	Sulfo-NHS-SS-Biotin 100 mg	100 mg
b2104-1gm	Sulfo-NHS-SS-Biotin 1 gram	1 gram
b2100-100mg	Sulfo-NHS-Biotin 100 mg	100 mg
b2100-1gm	Sulfo-NHS-Biotin 1 gram	1 gram
b2103-100mg	Sulfo-NHS-LC-Biotin 100 mg	100 mg
b2103-1gm	Sulfo-NHS-LC-Biotin 1 gram	1 gram
b2104-100mg	Sulfo-NHS-SS-Biotin 100 mg	100 mg
b2104-1gm	Sulfo-NHS-SS-Biotin 1 gram	1 gram
b2107-100mg	Biotin-HPDP 100 mg	100 mg
b2108-100mg	Iodoacetyl-LC-Biotin 100 mg	100 mg
b2106-100mg	Biotin Hydrazide 100 mg	100 mg
b2106-1gm	Biotin Hydrazide 1 gram	1 gram
b2111-100mg	D-Biotin 100 mg	100 mg