2. Others
  3. Fluorescent Dye
  4. 5-IAF

5-IAF  (Synonyms: 5-Iodoacetamidofluorescein)

Cat. No.: HY-D0807 Purity: ≥95.0%
COA Handling Instructions

Molarity Calculator

5-IAF (5-Iodoacetamidofluorescein) is an idoacetamide derivate of fluoresceine. 5-IAF can be used as fluorescent probe that labels proteins and other molecules having free thiols (cysteine side chains).

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

5-IAF Chemical Structure

5-IAF Chemical Structure

CAS No. : 63368-54-7

Size Price Stock Quantity
5 mg USD 84 In-stock
10 mg USD 134 In-stock
50 mg   Get quote  
100 mg   Get quote  

* Please select Quantity before adding items.

This product is a controlled substance and not for sale in your territory.

Customer Review

Based on 1 publication(s) in Google Scholar

5-IAF Related Antibodies

Top Publications Citing Use of Products

  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

5-IAF (5-Iodoacetamidofluorescein) is an idoacetamide derivate of fluoresceine. 5-IAF can be used as fluorescent probe that labels proteins and other molecules having free thiols (cysteine side chains)[1][2].

In Vitro

8 μM GSH sample solution is derivatized with 8, 80, 200, 400, 800, 1600, 2000, 2500 μM 5-IAF, respectively and analyzed with CE-LIF. The highest signal intensity is obtained with 400 and 800 μM 5-IAF[2].

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

5-IAF Related Antibodies
Molecular Weight

515.25

Formula

C22H14INO6
CAS No.
Appearance

Solid

Color

Light yellow to orange

Emission (Em)

515
Excitation (Ex)

488
SMILES

O=C(NC1=CC2=C(C3(C4=C(OC5=C3C=CC(O)=C5)C=C(O)C=C4)OC2=O)C=C1)CI
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 : 50 mg/mL (97.04 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 1.9408 mL 9.7040 mL 19.4081 mL
5 mM 0.3882 mL 1.9408 mL 3.8816 mL
View the Complete Stock Solution Preparation Table

* 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • 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% DMF    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (4.85 mM); Clear solution

  • Protocol 2

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

    Solubility: 2.5 mg/mL (4.85 mM); Suspended solution; Need ultrasonic

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: ≥95.0%

COA (232 KB)

Handling Instructions (2659 KB)
References
Kinase Assay
[2]

HepG2 cells are lysed in 1 mL 2% acetonitrile in water, and transfered to a 1.5 mL centrifuge tube. An additional 200 µL 2% acetonitrile is used to rinse the dish and combined with the cell extract. To measure the free GSH concentration, three aliquots of 80 µL cell extract are immediately transferred to 0.5 mL centrifuge tubes after a quick mix. Then 20 µL 50 µM NAC is added, followed by the addition of 100 µL 400 µM 5-IAF to the mixture. The derivatization is conducted at room temperature in dark for 1 h, after which the mixtures are centrifuged at 9000× g for 5 min to sediment the insoluble proteins. 10 µL supernatant is diluted 100× with water and analyzed immediately by CE-LIF[2].

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

Complete Stock Solution Preparation Table

* 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.9408 mL 9.7040 mL 19.4081 mL 48.5201 mL
5 mM 0.3882 mL 1.9408 mL 3.8816 mL 9.7040 mL
10 mM 0.1941 mL 0.9704 mL 1.9408 mL 4.8520 mL
15 mM 0.1294 mL 0.6469 mL 1.2939 mL 3.2347 mL
20 mM 0.0970 mL 0.4852 mL 0.9704 mL 2.4260 mL
25 mM 0.0776 mL 0.3882 mL 0.7763 mL 1.9408 mL
30 mM 0.0647 mL 0.3235 mL 0.6469 mL 1.6173 mL
40 mM 0.0485 mL 0.2426 mL 0.4852 mL 1.2130 mL
50 mM 0.0388 mL 0.1941 mL 0.3882 mL 0.9704 mL
60 mM 0.0323 mL 0.1617 mL 0.3235 mL 0.8087 mL
80 mM 0.0243 mL 0.1213 mL 0.2426 mL 0.6065 mL
  • No file chosen (Maximum size is: 1024 Kb)
  • If you have published this work, please enter the PubMed ID.
  • Your name will appear on the site.

5-IAF 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:

5-IAF63368-54-75-IodoacetamidofluoresceinFluorescent Dyefluorescent probecysteineprotein labelingInhibitorinhibitorinhibit

Your Recently Viewed Products:

Inquiry Online

Your information is safe with us. * Required Fields.

Product Name

  

Salutation

Applicant Name *

 

Email Address *

Phone Number *

 

Organization Name *

Department *

 

Requested quantity *

Country or Region *

      

Remarks

Bulk Inquiry

Inquiry Information

Product Name:
5-IAF
Cat. No.:
HY-D0807
Quantity:
MCE Japan Authorized Agent:

Customer Review

Thank You for Your Feedback!

Request for HNMR Report

Please fill out this form to request the QC report. We will send it to your Email address shortly.

Your information is safe with us. * Required Fields.

Product Name

  

Lot #

Applicant Name *

 

Email Address *

Phone Number

 

Department *

Organization Name *

 

Country or Region *

Remarks

Request for HNMR Report

We have received your request and will respond to you as soon as possible.