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  4. Vari Fluor 488 SE

Vari Fluor 488 SE  (Synonyms: VF 488 SE)

Cat. No.: HY-D1801 Purity: 75.80%
COA Handling Instructions

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Vari Fluor 488 SE (VF 488 SE) is a dye marker of the Vari Fluor SE series (Ex/Em=488 nm/513 nm). The Vari Fluor SE series of dyes are a class of fluorescent dyes containing NHS ester groups used to label free amines (-NHX) on antibodies, proteins, peptides, amine-modified oligonucleotides and other biomolecules.

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

Vari Fluor 488 SE

Vari Fluor 488 SE Chemical Structure

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1 mg USD 231 In-stock
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Vari Fluor 488 SE Related Antibodies

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Description

Vari Fluor 488 SE (VF 488 SE) is a dye marker of the Vari Fluor SE series (Ex/Em=488 nm/513 nm). The Vari Fluor SE series of dyes are a class of fluorescent dyes containing NHS ester groups used to label free amines (-NHX) on antibodies, proteins, peptides, amine-modified oligonucleotides and other biomolecules.

In Vitro

Protocol

1.Protein Preparetion
1) In order to obtain the best labeling effect, please prepare the protein (antibody) concentration as 2mg/mL.
2) The pH value of protein solution shall be 8.5±0.5. If the pH is lower than 8.0, 1 M sodium bicarbonate shall be used for adjustment.
3) If the protein concentration is lower than 2 mg/mL, the labeling efficiency will be greatly reduced. In order to obtain the best labeling efficiency, it is recommended that the final protein concentration range is 2-10 mg/mL.
4) The protein must be in the buffer without primary amine (such as Tris or glycine) and ammonium ion, otherwise the labeling efficiency will be affected.
2.Dye Preparation（Example for VF 488 SE）
Add anhydrous DMSO into the vial of VF 488 SE to make a 10 mg/mL stock solution. Mix well by pipetting or vortex.
3.Calculation of dye dosage
The amount of VF 488 SE required for reaction depends on the amount of protein to be labeled, and the optimal molar ratio of VF 488 SE to protein is about 10.
Example: assuming the required marker protein is 500 μL 2 mg/mL IgG (MW=150,000), use 100 μL DMSO dissolve 1 mg VF 488 SE, the required VF 488 SE volume is 5.05 μL, and the detailed calculation process is as follows:
1) mmol (IgG) = mg/mL (IgG)×mL (IgG) / MW (IgG) =2 mg/mL × 0.5 mL / 150,000 mg/mmol= 6.7×10-6 mmol
2) mmol (VF 488 SE) = mmol (IgG)× 10 = 6.7×10-6 mmol×10 = 6.7 × 10-5 mmol
3) μL (VF 488 SE) = mmol (VF 488 SE)×MW (VF 488 SE) / mg/μL (VF 488 SE) = 6.7 ×10-5 mmol ×834 mg/mmol / 0.01 mg/μL = 5.6 μL (VF 488 SE)
4.Run conjugation reaction
1) A good volume of freshly prepared 10 mg/mL VF 488 SE is slowly added to 0.5 mL protein sample.
In solution, gently shake to mix, then centrifuge briefly to collect the sample at the bottom of the reaction tube. Don't mix well to prevent protein samples from denaturation and inactivation.
2) The reaction tubules were placed in a dark place and incubated gently at room temperature for 60 minutes at intervals.For 10-15 minutes, gently reverse the reaction tubules several times to fully mix the two reactants and raise the bar efficiency.
5.Purify the conjugation
The following protocol is an example of dye-protein conjugate purification by using a Sephadex G-25 column.
1）Prepare Sephadex G-25 column according to the manufacture instruction.
2）Load the reaction mixture (From "Run conjugation reaction") to the top of the Sephadex G-25 column.
3）Add PBS (pH 7.2-7.4) as soon as the sample runs just below the top resin surface.
4）Add more PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate.

Note
1.CY dyes is sensitive to light and humidity. Immediately add CY dyesr solution and discard the unused part.
2.Low concentrations of sodium azide (≤3 mM or 0.02%) or thiomersal (≤0.02 mM or 0.01%) did not significantly interfere with protein labeling; However, 20-50% glycerol will reduce labeling efficiency.
3.Avoid buffering with primary amines (e.g., Tris, glycine) or ammonium ions,It compete with labeled proteins.
4.This product is only for scientific research by professionals, and shall not be used in clinical diagnosis or treatment, food or medicine.
5.For your safety and health, please wear lab coat and disposable gloves.

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

Vari Fluor 488 SE Related Antibodies
Appearance

Solid

Color

Orange to red

SMILES

[Vari Fluor 488 SE]
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

DMSO : 100 mg/mL (Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

  • 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: ≥ 2.5 mg/mL; Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 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: ≥ 2.5 mg/mL; Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 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).

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

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: 75.80%

COA (231 KB)

Handling Instructions (2659 KB)
References
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Vari Fluor 488 SE 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:

Vari Fluor 488 SEVF 488 SEFluorescent DyeVari Fluor SENHS esterlabeling free amineInhibitorinhibitorinhibit

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