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Biotin NHS  (Synonyms: Biotin N-hydroxysuccinimide ester; NHS-Biotin)

Cat. No.: HY-D0802 Purity: ≥98.0%
COA Handling Instructions

Molarity Calculator

Biotin NHS is an amino reactive biotin reagent used in the preparation of biotinylated surfaces or polypeptides.

For research use only. We do not sell to patients.

Biotin NHS Chemical Structure

Biotin NHS Chemical Structure

CAS No. : 35013-72-0

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Based on 10 publication(s) in Google Scholar

Biotin NHS Related Antibodies

Top Publications Citing Use of Products

  • Biological Activity

  • Purity & Documentation

  • References

  • Customer Review

Description

Biotin NHS is an amino reactive biotin reagent used in the preparation of biotinylated surfaces or polypeptides.

In Vitro

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
? Calculations:
? For best results, use ≥ 12-fold molar excess of biotin for a 10 mg/mL protein solution or ≥ 20-fold molar excess of biotin for a 2 mg/mL protein solution.
? 1.Calculate millimoles of biotin reagent to add to the reaction for a 20-fold molar excess:
? mL protein × (mg protein/mL protein) × (mmoL protein/mg protein) × (20 mmoL protein/mmoL protein) = mmoL Biotin.
? * 20 = Recommended molar fold excess of biotin for 2 mg/ml protein sample
? 2. Calculate microliters of 10 mM biotin reagent solution (prepared in Step B.3) to add to the reaction:
? mmoL Biotin × (1000000 μL/L) × (L/ 10 mmoL) =μL Biotin
? Biotin Labeling Reaction:
? 1. Dissolve 1-10 mg protein in 0.5-2.0 mL PBS according to the calculation made in Calculations.
? 2. Before use, prepare a 10 mM solution of the biotin reagent in an organic solvent such as DMSO.
? 3. Add the appropriate volume (μL) of 10 mM biotin reagent solution to the protein solution.
? 4. Incubate reaction on ice for two hours or at room temperature for 30 minutes.
? 5. To tests the labeled protein by ELISA or Western blot.

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

Biotin NHS Related Antibodies
Molecular Weight

341.38

Formula

C14H19N3O5S
CAS No.
Appearance

Solid

Color

White to off-white

SMILES

O=C(ON1C(CCC1=O)=O)CCCC[C@@H]2SC[C@]([C@]2([H])N3)([H])NC3=O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

-20°C, sealed storage, away from moisture and light

*The compound is unstable in solutions, freshly prepared is recommended.

Solvent & Solubility
In Vitro: 

DMSO : 62.5 mg/mL (183.08 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.9293 mL 14.6464 mL 29.2929 mL
5 mM 0.5859 mL 2.9293 mL 5.8586 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.08 mg/mL (6.09 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.08 mg/mL (6.09 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*The compound is unstable in solutions, freshly prepared is recommended.

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: ≥98.0%

COA (249 KB) HNMR (253 KB) OR (712 KB)

Handling Instructions (2659 KB)
Dyeing Example
References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 2.9293 mL 14.6464 mL 29.2929 mL 73.2322 mL
5 mM 0.5859 mL 2.9293 mL 5.8586 mL 14.6464 mL
10 mM 0.2929 mL 1.4646 mL 2.9293 mL 7.3232 mL
15 mM 0.1953 mL 0.9764 mL 1.9529 mL 4.8821 mL
20 mM 0.1465 mL 0.7323 mL 1.4646 mL 3.6616 mL
25 mM 0.1172 mL 0.5859 mL 1.1717 mL 2.9293 mL
30 mM 0.0976 mL 0.4882 mL 0.9764 mL 2.4411 mL
40 mM 0.0732 mL 0.3662 mL 0.7323 mL 1.8308 mL
50 mM 0.0586 mL 0.2929 mL 0.5859 mL 1.4646 mL
60 mM 0.0488 mL 0.2441 mL 0.4882 mL 1.2205 mL
80 mM 0.0366 mL 0.1831 mL 0.3662 mL 0.9154 mL
100 mM 0.0293 mL 0.1465 mL 0.2929 mL 0.7323 mL
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Biotin NHS Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Biotin NHS35013-72-0Biotin N-hydroxysuccinimide ester NHS-BiotinFluorescent DyeInhibitorinhibitorinhibit

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