Anti-Sml1 | Suppressor of Mec1 lethality, Rabbit, Polyclonal
Artikelnummer:
AGR-AS10-847
Hersteller Artikelnummer:
AS10-847
Alternativnummer:
AGR-AS10-847
Hersteller:
Agrisera
Wirt:
Rabbit
Kategorie:
Antikörper
Applikation:
WB
Spezies Reaktivität:
Yeast
Immunogen:
KLH-conjugated synthetic peptide derived from known S.cerevisie Sml1 sequence. Gene ID: 854945
Sml1 is a ribonucleotide reductase inhibitor involved in regulation of dNTP production. It is regulated by Mec1p and Rad53p during DNA damage and S phase. Synonymes: Yml058w.
Cell preparation for western blot: cells were harvested by centrifugation (4000 x g , 6 min, 4C). Supernatant was discarded and cells were resuspended in 500 µl cold TCA buffer (20 mM Tris, pH 8, 50 mM ammonium acetate, 2 mM EDTA, 1 tablet/10 ml of Complete Mini Protease inhibitor cocktail with EDTA (Roche Diagnostics GmbH)). 500 µl 0.5 mm Zirconia/Silica Beads (BioSpec Products, Inc, 11079105z) and 500 µl cold 20% TCA was added. Samples were vigorously vortexed twice for 30 s (kept on ice in between), 750 µl from the liquid phase was transferred into a fresh Eppendorf tube. Samples were centrifuged for 10 min (20000 x g, 4C). The pellet was resuspended in 300 µl TCA-Laemmli buffer and boiled for 10 min at 100C.
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