MYC-Nanoab-Magnetic Beads

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Protein Biology

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La description

MYC-Nanoab-Magnetic Beads CATï¼šMNM-25-1000

product ID: MNM-25-1000
Storage conditions:Stored at -20â„ƒ for one year(to ensure full sealing), avoid centrifugation, drying and freeze-thaw.

Product description
Magnetic nanospheres of coupling anti-Myc-tag nanoantibodies are used for immunoprecipitation of Myc-tag (EQKLISEEDL) fusion protein
Product advantages
No heavy chain and light chain, clean background;
Ready-to use type, saving time;
High affinity and high binding ability.
Scope of application
It can be used for immunoprecipipation(IP)/immunocoprecipipation(CoIP)ã€chromatin immunoprecipitation(ChIP)/RNA-binding protein immunoprecipipation(RIP), enzyme activity determination, mass spectrometry analysis, etc.
Specificity
The specific binding is located at the N-terminal, C-terminal and internal Myc-tag of the fusion protein. Does not bind to endogenous c-MYC.
Product features
Storage buffer: PBS(containing 20% ethanol);
Storage conditions: Stored at 4â„ƒ for one year(to ensure full sealing), avoid centrifugation, drying and freeze-thaw.
Experimental principle

Experimental steps:
Plant tissue cracking treatment:
Take an appropriate amount of plant tissue samples (leaves, etc.) and place them in frozen liquid nitrogen, and then put the frozen plant tissue samples in a mortar for grinding, so as to fully grind and destroy their cell walls. Add 500-1000ul RIPA lysate (added with protease inhibitor PMSF, etc.) for cracking. In order to improve the cracking efficiency, add 200ul glass powder and fully shake for 30min. After cracking, 12000 rpm, centrifuge for 30min, suck the supernatant and place it in a new centrifuge tube, and discard the sediment.
Collect cells
Approximately 106-107 cells expressing Myc-tag fusion protein were used for each immunoprecipitation reaction, and the number of cells could be appropriately adjusted according to the expression of Myc-tag fusion protein. Suck out the medium and add precooled 1 × PBS, rinse twice, collect adherent cells by cell scraping or trypsin digestion, transfer the cells to a centrifuge tube, centrifuge 500 g for 3 minutes and discard the supernatant.

Cell lysis
1. Protease inhibitors are added to the lysis buffer, and the cells are suspended with a pre-cooled lysis buffer of 500μl.
2. Put it on the ice for 30 minutes and blow it fully every 10 minutes.
3.Centrifuge 20000 g at 4 â„ƒ for 15 minutes, transfer the cracking product (supernatant) to a new precooling tube, and discard the precipitation.
Note: At this time, the cell lysis product can be preserved at -80°C for a long time.
Balanced beads
4. Fully mix Myc-Nanoab-Magnetic Beads, absorb 40μl of the product into the 500μl precooled cracking buffer, separate the magnetic beads on the magnetic frame until the top becomes clear, and discard the liquid.( This step is optional)
Binding protein
5. The balanced Myc-Nanoab-Magnetic Beads are added to the cell lysis product (40μl can be added directly to the cell lysis product if step 4 is not done) and rotate and combine at 4°C for 1 hour. The combination time can be adjusted according to the needs of the experiment. If necessary, the 50μl cracking product is retained for immunoprinting analysis.
6. Separate the magnetic beads on the magnetic shelf until the upper clear becomes clear and discard the upper liquid. If necessary, 50μl supernatant is retained for immunoprinting analysis.
Wash the beads
7. Use 500 μL precooled pyro lysis buffer resuspended Myc-Nanoab-Magnetic Beads, separated the magnetic beads on the magnetic shelf until the supernatant became clear, discarded the supernatant and washed again for 3 times. Minimize cleaning time.
Elution protein
Method 1:
8. Adding 20μl 2×SDS-sample buffer suspends Myc-Nanoab-Magnetic Beads.95â„ƒ, heated for 10 min, fully denatured, separated on the magnetic shelf and the collected products can be analyzed by SDS-PAGE and immunoblotting.

Pays:	China
N Â° de modÃ¨le:	-
Prix FOB:	(Negotiable) Obtenir le dernier prix
LocalitÃ©:	China
Prix de commande minimale:	-
Commande minimale:	-
Packaging Detail:	-
Heure de livraison:	7-15 days
Capacité de Fournir:	-
Payment Type:	T/T, L/C, D/A, D/P, Western Union, Money Gram, PayPal, Other
Groupe de produits :	Protein Biology