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THP-1 (human acute monocytic leukemia) Nuclear extract lysate, Denatured; Abnova

Artikelnummer. 16019515
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Mængde:
50 μg
Pakningsstørrelse:
50µg
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16019515 50 μg 50µg
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Artikelnummer. 16019515 Leverandør Abnova Leverandørnr. L007V3.50ug

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Western Blotting

Tekniske data

Til brug med (applikation) Western Blot
Væv Blood, peripheral
Oversigt Nuclear extract cell lysate (denatured)
Lyseringsbuffer Buffer A: 10mM HEPES pH 7.9, 1.5mM MgCl2, 10mM KCl, 0.5 mM DTT. Buffer C: 20mM HEPES pH 7.9, 25%(v/v) Glycerol , 0.42M NaCl , 1.5mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT & 0.5 mM PMSF. Buffer D : 20mM HEPES pH 7.9, 20%(v/v) glycerol, 50mM KCl, 0.2 mM EDTA, 0.5 mM DTT & 0.5 mM PMSF.
Fremstillingsmetode Nuclear extract was prepared by using a modified protocol of Dignam et al. Cells were Harvested and homogenized in Buffer A, and then centrifugated at 25,000 g for 20 minutes to remove cytoplasm and pellet the nuclei. The pellet was re-suspended in Buffer C, and then the suspensions were centrifuged to collect nuclear extract. The supernatant was dialyzed against Buffer D. The dialysate was then centrifuged, divided into aliquots, and stored at -80°C. The protein concentration was determined by the method of Bradford (Bio-Rad protein assay, microplate standard assay). The lysate was adjusted to 2.5 mg/ml, and then mixed with 5X Sample Buffer to become final 2 mg/ml in 1X Sample Buffer. The lysate was heated at 95°C for 5 min, and cooled rapidly.
Kvalitetskontrol test 12.5% SDS-PAGE Stained with Coomassie Blue.
Mængde 50 μg
Opbevaringsbuffer In ready-to-use 1X Sample Buffer (50 mM Tris-HCl, 2% SDS, 10% glycerol, 300 mM 2-mercaptoethanol, 0.01% Bromophenol blue).
Værtsarter Human
Koncentration 2 mg/mL
Indhold og opbevaring Store at -80°C. Aliquot to avoid repeated freezing and thawing.
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