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  5. 3X FLAG peptide TFA

3X FLAG peptide TFA is a FLAG-tagged peptide containing three repeats of the Asp-Tyr-Lys-Xaa-Xaa-Asp motif. 3X FLAG peptide TFA can be used for protein separation and purification, and competitive elution with target proteins.

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3X FLAG peptide TFA Chemical Structure

3X FLAG peptide TFA Chemical Structure

Size Price Stock Quantity
1 mg USD 72 In-stock
5 mg USD 132 In-stock
10 mg USD 228 In-stock
25 mg USD 504 In-stock
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Customer Review

Based on 8 publication(s) in Google Scholar

Other Forms of 3X FLAG peptide TFA:

Top Publications Citing Use of Products
  • Biological Activity

  • Purity & Documentation

  • References

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Description

3X FLAG peptide TFA is a FLAG-tagged peptide containing three repeats of the Asp-Tyr-Lys-Xaa-Xaa-Asp motif. 3X FLAG peptide TFA can be used for protein separation and purification, and competitive elution with target proteins.

In Vitro

1. Purification of FLAG tagged proteins through 3X FLAG peptide competition[2]
(1) Plasmid transfection: Transfect plasmids of mIRS-1, hIRS-1, or their mutants labeled with FLAG into 293 T cells for 2 days.
(2) Cell collection: Cells were harvested by centrifugation at 14000 rpm for 10 minutes at 4 °C, and lysed with HEPES buffer (150 mM NaCl, 50 mM HEPES, pH 7.4, 5 mM EDTA, 1% (w/v) sodium deoxycholate, 1% (v/v) Triton X-100, 0.2% sodium fluoride, 0.1% sodium orthovanadate, and protease inhibitor mixture).
(3) Using Anti-FLAG magnetic beads for protein separation and purification: Add Anti-Flag magnetic beads (HY-K0207)/Anti-c-Myc magnetic beads (1 μm) (HY-K0207A) to the cell lysate, and incubate at room temperature for 2 hours or at 4 °C overnight. Then wash with a washing buffer and elute with a buffer containing 1 mg/mL 3X FLAG. Finally, use a gel filtration column for further purification of the supernatant, which is equilibrated with the storage buffer. All purified proteins are concentrated using centrifugal filtration and aliquoted for storage at -80 °C.
(4) Protein quantification and validation: The purified protein was quantified using an ND-2000 NanoDrop spectrophotometer with OD 280 and validated by Coomassie Brilliant Blue staining.
2. Co-IP and co-IP-MS analysis using the 3X FLAG peptide[3]
(1) Centrifuge the cell lysate at 13300 rpm for 15 minutes at 4 °C to remove intact cells.
(2) The supernatant was incubated overnight with control immunoglobulin G (IgG) or primary antibody in immunoprecipitation buffer (150 mM NaCl, 20 mM HEPES, pH 7.4, 1% Triton X-100, 12.5 mM β - glycerophosphate, 1.5 mM MgCl2, 2 mM EGTA, containing phosphatase and protease inhibitors), and then incubated with 20 μL resuspended A/G protein beads at 4 °C for 2 hours to precipitate bound proteins.
(3) For continuous immunoprecipitation, incubate 3X FLAG eluent solution (final concentration 150 ng/mL) with 5 times the volume of gel at 4 °C for 2 hours, elute the bound protein from beads, and then use secondary antibody immunoprecipitation at 4 °C overnight. The beads were centrifuged at 1000 g for 5 minutes at 4 °C to remove the supernatant, washed 4 times with immunoprecipitation buffer, and boiled at 95 °C for 20 minutes.
(4) The samples were run on SDS-PAGE gel and then dyed (Bio Rad). Subsequently, cut off the tape, decolorize, digest with trypsin, and perform MS analysis to determine individual proteins using liquid chromatography-MS.

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

Molecular Weight

2975.84

Formula

C122H170F3N31O51S

Appearance

Solid

Color

White to off-white

Sequence

Met-Asp-Tyr-Lys-Asp-His-Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys

Sequence Shortening

MDYKDHDGDYKDHDIDYKDDDDK

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Sealed storage, away from moisture

Powder -80°C 2 years
-20°C 1 year

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

Solvent & Solubility
In Vitro: 

H2O : 25 mg/mL (8.40 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 0.3360 mL 1.6802 mL 3.3604 mL
5 mM 0.0672 mL 0.3360 mL 0.6721 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 (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • 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:

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
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:  PBS

    Solubility: 50 mg/mL (16.80 mM); Clear solution; Need ultrasonic and adjust pH to 3 with HCl

  • Protocol 2

    Add each solvent one by one:  20 mM Tris pH7.5, 100 mM NaCl

    Solubility: 33.33 mg/mL (11.20 mM); Clear 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.
Calculation results:
Working solution concentration: mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
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).
Purity & Documentation

Purity: 99.79%

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 (sealed storage, away from moisture). 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
H2O 1 mM 0.3360 mL 1.6802 mL 3.3604 mL 8.4010 mL
5 mM 0.0672 mL 0.3360 mL 0.6721 mL 1.6802 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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3X FLAG peptide TFA 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.

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Product Name:
3X FLAG peptide TFA
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HY-P0319A
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