TAMRA-Azide-PEG-biotin 

TAMRA-Azide-PEG-biotin is a dye derivative of TAMRA (HY-135640). TAMRA-Azide-PEG-biotin contains an Azide group and can undergo copper-catalyzed azide-alkyne cycloaddition reaction (CuAAc) with molecules containing Alkyne groups. It can also undergo strain-promoted alkyne-azide cycloaddition (SPAAC) reactions with molecules containing DBCO or BCN groups.

Guidelines (The following is our recommended protocol. This protocol is provided for reference only and should be modified according to your specific needs).
Enrichment of N-myristoylated proteins^[1]
1. 293T cells are treated with 40 μM Alkynyl myristic acid (YnMyr) (HY-140335) for 24 h followed by lysis in PBS containing 1% Triton X-100, 0.1% SDS and protease inhibitors.
2. Proteins are diluted to 1 mg/mL with lysis buffer.
3. 100 μg of proteins are used to perform click chemistry.
4. A click mixture is prepared by adding reagents in the following order and by vortexing between the addition of each reagent:
1 μL of 10 mM capture reagent (TAMRA-Azide-PEG-biotin) (dissolved in DMSO), 2 μL of 50 mM CuSO₄, 2 μL of 50 mM TCEP (HY-W011500), 1 μL of 10 mM Tris(benzyltriazolylmethyl)amine (TBTA) (HY-116677).
5. 6 μL of the click reagent mixture is added into each protein sample followed by incubation for 1 h at room temperature with gentle votexing.
6. The click reaction is halted with 5 mM EDTA (HY-Y0682), and proteins are precipitated by adding 200 μL water, 400 μL methanol and 100 μL chloroform followed by centrifuging at 6,000 × g for 5 min.
7. After washing once with methanol, the precipitated proteins are dissolved in 10 μL PBS containing 2% SDS and 10 mM DTT (HY-15917) followed by diluted 10-fold with PBS.
8. For the affinity enrichment, the proteins are incubated with 30 μL Streptavidin Magnetic Beads (HY-K0208) for 2 h at room temperature with gentle mixing.
9. After incubation, the supernatant is collected as the unbound fraction and the beads washed 5 times with PBS containing 0.2% SDS.
10. Then the proteins are resolved by SDS-PAGE and in-gel fluorescence is scanned at 532/575 nm excitation/emission wavelengths using fully automatic luminescence imaging system followed by immunoblot.

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

1174.38

C₅₇H₇₉N₁₁O₁₄S

1797415-74-7

Solid

Purple to purplish red

CN(C)C1=CC([O+]=C(C=C(N(C)C)C=C2)C2=C3C4=CC=C(C(NCCC(N(CCC(NCCOCCOCCOCCNC(CCCCC5SC(NC(N6)=O)C6C5)=O)=O)CCOCCOCCOCCN=[N+]=[N-])=O)=O)C=C4C([O-])=O)=C3C=C1.CN(C)C7=CC([O+]=C(C=C(N(C)C)C=C8)C8=C9C%10=CC(C(NCCC(N(CCC(NCCOCCOCCOCCNC(CCCCC%11SC(NC(N%12)=O)C%12C%11)=O)=O)CCOCCOCCOCCN=[N+]=[N-])=O)=O)=CC=C%10C([O-])=O)=C9C=C7

Room temperature in continental US; may vary elsewhere.

* 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. 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.

• [1]. An S, et al. Small-molecule PROTACs: An emerging and promising approach for the development of targeted therapy drugs. EBioMedicine. 2018 Oct;36:553-562  [Content Brief]

  [2]. Zhang X, et al. CRISPR screening identifies PRMT1 as a key pro-ferroptotic gene via a two-layer regulatory mechanism. Cell Rep. 2024 Sep 24;43(9):114662.  [Content Brief]