Fig. 1 · Structure

Cat. No.: A44814
CAS No.: 191671-46-2
Formula: C₂₀H₂₉N₄NaO₉S₂
Mol. Weight: 556.59

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Form	MW
Sulfo-NHS-LC-Biotin sodium This page	556.59
Sulfo-NHS-LC-Biotin	534.60

Sulfo-NHS-LC-Biotin sodium

Amine-reactive Reagent

Sulfo-NHS-LC-Biotin sodium is an amine-reactive ester derivative of biotin that primarily targets amine groups for covalent labeling. Due to its inability to cross the intact blood-retinal barrier, it serves as a valuable tool in assessing vascular permeability within the brain and retinal vasculature. This reagent is beneficial for studies investigating neurovascular dynamics and the integrity of blood-brain and blood-retinal barriers.

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Biological Activity

Bioactivity / Mechanism

Sulfo-NHS-LC-Biotin is used to biotinylate cells before avidin-mediated attachment of biotinylated cargo. In C3H10T1/2 cells, increasing the labeling concentration increased cell-associated NanoLuc activity. [1]

Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.

In Vitro Studies

In C3H10T1/2 cells, viability was largely maintained at labeling concentrations up to 1000 μM, while 5000 and 10000 μM reduced viability. These findings apply to the reported surface-modification conditions. [1]

Concentration-dependent cell biotinylation and reporter attachment

Cell-based study · Murine C3H10T1/2 mesenchymal stem cell line [1]

The concentration-response design assessed viability and avidin-mediated NanoLuc attachment. Labeling increased over the tested reporter-attachment range; the highest concentrations in the viability panel caused a modest significant reduction. These are labeling concentrations, not potency estimates.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Cell viability following surface modification, measured 24 h after seeding	CCK-8 viability Hardly affected at 1000 μM or lower; significantly reduced at 5000 and 10000 μM (Figure 2A, *p < 0.05 versus control).	[1]
Cell-associated NanoLuc activity after avidin/biotin-NanoLuc attachment	RLU / 10^5 cells 1000 μM produced higher reporter attachment than 500 μM; exact bar heights were not extracted.	[1]

Literature-Reported Treatment Conditions

Treatment concentration: Viability: 50, 100, 500, 1000, 5000 or 10000 μM; reporter attachment: 50, 100, 500 or 1000 μM.
Exposure duration: Biotinylation: 10 min at room temperature; viability assessed 24 h after seeding surface-modified cells.
Method: Cell-surface biotinylation followed by washing and avidin-mediated reporter attachment.
Vehicle: HBSS
Co-treatment: Avidin 100 μg/mL for 5 min, followed by biotin-NanoLuc 100 μM for 5 min.
Reported limits: No analytical characterization of the sodium form or stereochemical purity. Exact viability percentages and reporter bar heights are not tabulated in the selected locations.

Methods and source locations

Compound reported in the paper: Sulfo-NHS-LC-biotin; Compound label in the paper: Pierce sulfosuccinimidyl-6-[biotin-amido]hexanoate

Research use of the named commercial reagent for cell biotinylation; no independent sodium-form comparison or Adooq validation.

Figure 2A–B and caption; Results: Optimisation of biotinylation of cells and of surface modification; Methods: Materials; Surface modification of cells by the ABC method

Cell viability following surface modification, measured 24 h after seeding: CCK-8; Figure 2 caption states experiments conducted in duplicate, representative experiment shown, results mean ± SD of three to four experiments. This replication wording is retained without reinterpretation. — Figure 2A and caption; Methods: Surface modification of cells by the ABC method

Cell-associated NanoLuc activity after avidin/biotin-NanoLuc attachment: Nano-Glo luciferase assay in lysed cells; concentration-dependent comparison in Figure 2B. Same caption replication wording as Figure 2A. — Figure 2B and caption; Results: Optimisation of biotinylation of cells and of surface modification; Methods: Surface modification of cells by the ABC method

In Vivo Studies

Sulfo-NHS-LC-Biotin served as a perfused vascular tracer in P15 mice. Labeling remained within control retinal vessels but was occasionally extravascular in Eogt-deficient retinas, demonstrating a barrier-assessment application. [2]

Retinal vascular permeability tracing in P15 mice

Animal study · Postnatal day 15 wild-type and Eogt−/− mouse retinas [2]

Perfused reagent was used as a vascular tracer. Streptavidin-detected biotin remained intravascular in control retinas but was occasionally present outside vessels in Eogt-deficient retinas. The genetic model determines barrier integrity; the reagent is the readout.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Localization of perfused biotin tracer relative to IB4-positive retinal vessels	Qualitative extravasation Control P15 retinas: labeling remained within vessels; Eogt−/− retinas: occasional extravascular labeling.	[2]

Conditions shared by the study groups

Route: Perfusion (anatomical entry site not specified in the primary methods subsection)
Frequency: Single perfusion
Treatment course: 10 min; immediate post-perfusion assessment
Vehicle: 10 ml PBS-CMF

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
Wild-type P15 mice 0.75 μg/g mouse weight	Wild-type retinas receiving the same tracer perfusion	Biotin labeling remained within retinal vessels.	[2]
Eogt−/− P15 mice 0.75 μg/g mouse weight	Wild-type retinas receiving the same tracer perfusion	Biotin labeling was occasionally detected in extravascular spaces.	[2]

Wild-type P15 mice: Model: Wild-type P15 mice; Reported limits: Mouse sex and total mouse n are not specified in the selected panel-C subsection.

Eogt−/− P15 mice: Model: Eogt−/− P15 mice; Reported limits: Mouse sex and total mouse n are not specified in the selected panel-C subsection.

Methods and source locations

Compound reported in the paper: Sulfo-NHS-LC-biotin; Compound label in the paper: Thermo Fisher Scientific 21335

Tracer application of the named reagent; no Adooq lot validation or pharmacological efficacy claim.

Figure 7C and caption; Results: EOGT is required for retinal vascular integrity; Methods: Sulfo-NHS-LC-biotin perfusion in retina

Localization of perfused biotin tracer relative to IB4-positive retinal vessels: CF488A-conjugated streptavidin with Dylight594-IB4 and confocal microscopy; Figure 7C maximum-intensity projections. No leakage counts or mouse n inferred from images. — Figure 7C and caption; Results: EOGT is required for retinal vascular integrity; Methods: Sulfo-NHS-LC-biotin perfusion in retina

Wild-type P15 mice · Figure 7C and caption; Results: EOGT is required for retinal vascular integrity; Methods: Sulfo-NHS-LC-biotin perfusion in retina

Eogt−/− P15 mice · Figure 7C and caption; Results: EOGT is required for retinal vascular integrity; Methods: Sulfo-NHS-LC-biotin perfusion in retina

Specifications

Manufacturer	Adooq Bioscience
CAS Number	191671-46-2
Applications	Amine-reactive Reagent
Formula	C₂₀H₂₉N₄NaO₉S₂
Molecular Weight	556.59
SMILES	O=C(C(CC1=O)S(=O)(O[Na])=O)N1OC(CCCCCNC(CCCC[C@H]2[C@]3([H])[C@](NC(N3)=O)([H])CS2)=O)=O

Storage & Solubility

In Vitro Solubility

Reported Solubility

No reported solubility data are available for this product.

Preparing Stock Solutions

Stock preparation is unavailable for this product molecular weight.

Calculated stock concentrations do not establish solvent solubility. Refer to the reported solubility data for experimental guidance.

Storage

Store powder at -20°C, keep desiccated.
In powder form, the chemical is stable for 36 months.
In solution, store at -20°C and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Research References

[1] Takayama Y, Kusamori K, Hayashi M, et al. Long-term drug modification to the surface of mesenchymal stem cells by the avidin-biotin complex method. Scientific reports (2017). PMID: 29208980 DOI: 10.1038/s41598-017-17166-8 Source material
[2] Sawaguchi S, Varshney S, Ogawa M, et al. O-GlcNAc on NOTCH1 EGF repeats regulates ligand-induced Notch signaling and vascular development in mammals. eLife (2017). PMID: 28395734 DOI: 10.7554/elife.24419 Source material

Publications

1 publication citing this product

Dongdong Xu, .et al. , Chem Sci, 2020, 11, 4422-4429

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Sulfo-NHS-LC-Biotin sodium

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