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SpCas9 (D10A/H840A) 3NLS

recombinant, E. coli

dead SpCas9 (SpdCas9)

Cat. No. Amount Price (EUR) Buy / Note
RNT-011-S 10 μl (20 μM) 125,00 Add to Basket/Quote Add to Notepad
RNT-011-L 5x 10 μl (20 μM) 495,00 Add to Basket/Quote Add to Notepad

For in vitro use only!

Shipping: shipped on blue ice

Storage Conditions: store at -20 °C
avoid freeze/thaw cycles

Shelf Life: 12 months

Purity: ≥ 95 % (SDS-PAGE)

Form: liquid

Concentration: 20 μM

Description:
CRISPR-associated (Cas) 9 protein from Streptococcus pyogenes is an RNA-guided endonuclease suitable for ribonucleoprotein (RNP) complex formation with SpCas9-specific sgRNA for subsequent gene targeting both in vitro and in living cells[1]. The wild-type SpCas9 possesses two conserved nuclease domains: HNH-like and RuvC-like that generate double-strand breaks by cleaving the complementary DNA strand and opposite strand, respectively.[1]

SpCas9 (D10A/H840) 3NLS contains mutations within the RuvC-like (D10A) and HNH-like (H840A) nuclease domains that inactivate cleavage activity but still allow RNA-guided sequence-specific DNA binding1. It can therefore be used as negative control or visualiztation tool in combination with labeled target specific sgRNA.

SpCas9 (D10A/H840A) 3NLS has an apparent molecular weight of approx. 160 kDa (SDS-PAGE) and contains a C-terminal 6x His-tag as well as three C-terminal nuclear localization sequences (NLS) for improved nuclear localization.

Content:
SpCas9 (D10A/H840A) 3NLS
#RNT-010-S: 1x 10 μl (20 μM)
#RNT-010-L: 5x 10 μl (20 μM)
50 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, 1 mM EDTA, 50% Glycerol (v/v)

SpCas9 Reaction Buffer
1x 1 ml (10x)
200 mM HEPES (pH 6.5), 50 mM MgCl2, 1 M NaCl, 1 mM EDTA

SpCas9 Dilution Buffer
1x 1 ml
50 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, 1 mM EDTA, 50% Glycerol (v/v)

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Selected References:
[1] Jinek et al. (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337(6096):816.