Revision 3
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC

REACTIVITY:

All

SENSITIVITY:

Transfected Only

MW (kDa):

143

Source/Isotype:

Rabbit IgG

UniProt ID:

#A0A182DWE3

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

LbCpf1/Cas12a (Strain ND2006) (E8B1W) Rabbit mAb recognizes transfected levels of total LbCpf1/Cas12a (Strain ND2006) protein. This antibody does not cross-react with Cas9 (S. pyogenes), Cas9 (S. aureus), AsCpf1/Cas12a (Strain BV3L6), or FnCpf1/Cas12a (Strain U112) protein.

Species Reactivity:

All Species Expected

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of LbCpf1/Cas12a (Strain ND2006) protein.

Background

CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins) are RNA-guided nuclease effectors that are utilized for precise genome editing in mammalian systems (1). Cpf1/Cas12a (CRISPR from Prevotella and Francisella) proteins are members of the Class 2 CRISPR system (2). Class 2 CRISPR systems, such as the well characterized Cas9, rely on single-component effector proteins to mediate DNA interference (3). Cpf1/Cas12a endonucleases, compared to Cas9 systems, have several unique features that increase the utility of CRISPR-based genome editing techniques: 1) Cpf1/Cas12a-mediated cleavage relies on a single and short CRISPR RNA (crRNA) without the requirement of a trans-activating crRNA (tracrRNA), 2) Cpf1/Cas12a utilizes T-Rich protospacer-adjacent motif (PAM) sequences rather than a G-Rich PAM, and 3) Cpf1/Cas12a generates a staggered, rather than a blunt-ended, DNA double-stranded break (2). These features broaden the utility of using CRISPR-Cas systems for specific gene regulation and therapeutic applications. Several Cpf1/Cas12a bacterial orthologs have been characterized for CRISPR-mediated mammalian genome editing (2,4).~LbCpf1 (Strain ND2006)/Cas12a is a Cpf1/Cas12a enzyme derived from Lachnospiraceae bacterium ND2006 (5,6).

  1. Cong, L. et al. (2013) Science 339, 819-23.
  2. Zetsche, B. et al. (2015) Cell 163, 759-71.
  3. Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
  4. Zhang, Y. et al. (2017) Sci Adv 3, e1602814.
  5. Zetsche, B. et al. (2015) Cell 163, 759-71.
  6. Zhang, Y. et al. (2017) Sci Adv 3, e1602814.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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