Sulfo-NHS-LC-LC-Biotin  (Synonyms: Biotin-XX-SSE)

Sulfo-NHS-LC-LC-Biotin (Biotin-XX-SSE), a biotin reagent, is used to label the proteins exposed to the external leaflet of intact exosomes and contains a larger spacer arm between the biotin and amine reactive linker. The size of this linker helps to overcome steric hindrance and increases labeling efficiency at the crowded exosome surface^[1][2].

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
Calculation: To achieve the best results, use a biotin concentration of ≥ 12 times molar excess for the 10 mg/mL protein solution, or ≥ 20 times molar excess for the 2 mg/mL protein solution.
Calculate the number of millimoles of biotin reagent to be added to the reaction to achieve a 20-fold molar excess:
mL of protein × (mg of protein/mL of protein) × (mmol of protein/mg of protein) × (20 mmol of protein/mmol of protein) = mmoL of biotin.
20 = The recommended molar excess of biotin for a 2 mg/ml protein sample.
2. The volume (in microliters) of the 10 mM biotin solution to be added to the reaction:
mmoL biotin × (1000000 μL/L) × (L/10 mmoL) = μL biotin Preparation of Biocytin-Linked Antibody 1 (bio-Ab1)^[1] Add 200 μL of the antibody (0.83 μM, dissolved in PBS) and 1.66 μL of 2 mM Sulfo-NHS-LC-LC-Biotin (dissolved in DMSO) to a reaction vessel, and let it react at room temperature for 1 hour with continuous shaking.
2. The reaction mixture was subjected to buffer exchange and then purified using a Zeba rotary desalination column (with a molecular weight cut-off of 40K, balanced with PBS) to remove the unbound biotin.
3. Stabilize the purified biotinylated antibody (bio-Ab1) in PBS buffer containing 0.1% (w/v) BSA, and store it at 4℃.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

669.74

C₂₆H₄₀N₅NaO₁₀S₂

194041-66-2

Solid

White to off-white

O=C(C(CC1=O)S(=O)(O[Na])=O)N1OC(CCCCCNC(CCCCCNC(CCCC[C@H]2[C@]3([H])[C@](NC(N3)=O)([H])CS2)=O)=O)=O

Room temperature in continental US; may vary elsewhere.

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Diaz G, et al. Changes in the Membrane-Associated Proteins of Exosomes Released from Human Macrophages after Mycobacterium tuberculosis Infection. Sci Rep. 2016 Nov 29;6:37975.  [Content Brief]

  [2]. Gabant G, et al. Assessment of solvent residues accessibility using three Sulfo-NHS-biotin reagents in parallel: application to footprint changes of a methyltransferase upon binding its substrate. J Mass Spectrom. 2008 Mar;43(3):360-70.  [Content Brief]

  [3]. Ilia Fishbein, et al. Post-Deployment Modifications of Stent with Endothelial Cells. CARDIOVASCULAR AND PULMONARY DISEASES, 24, SUPPLEMENT 1, S68, MAY 01, 2016.

  [4]. Lei H, Tang Y. A novel preparation method for monodisperse streptavidin magnetic beads and its application in electrochemiluminescent immunoassay for CEA. PLoS One. 2025 Dec 9;20(12):e0338214.  [Content Brief]