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Key Documents

MAC148

Sigma-Aldrich

Anti-Cas9 Antibody, clone 8G4

Synonyme(s) :

CRISPR-associated endonuclease Cas9/Csn1, SpCas9, SpyCas9

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About This Item

Code UNSPSC :
12352203
Nomenclature NACRES :
NA.43

Source biologique

rat

Niveau de qualité

Forme d'anticorps

purified antibody

Type de produit anticorps

primary antibodies

Clone

8G4, monoclonal

Poids mol.

calculated mol wt 158.44 kDa
observed mol wt ~210 kDa

Produit purifié par

using protein G

Espèces réactives

all

Conditionnement

antibody small pack of 100

Technique(s)

immunofluorescence: suitable
western blot: suitable

Isotype

IgG2aλ

Séquence de l'épitope

Unknown

Numéro d'accès Protein ID

Numéro d'accès UniProt

Température de stockage

2-8°C

Informations sur le gène

Streptococcus pyogenes ... Cas9(69900935)

Spécificité

Clone 8G4 is a rat monoclonal antibody that specifically detects Cas9.

Immunogène

His-tagged, full-length, recombinant Cas9 from Streptococcus pyogenes.

Application

Quality Control Testing

Evaluated by Western Blotting in lysate from HeLa cells overexpressing Cas9.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Cas9 in lysate from HeLa cells overexpressing Cas9.

Tested Applications

Immunofluorescence Analysis: A 1:500 dilution from a representative lot detected Cas9 in Cell line stably expressing Cas9-NLS (A) versus wild type parental cell line (B) (Courtesy of Germán Camargo Ortega (Institute of Stem Cell Research, Helmholtz Munich,. Germany).

Western Blotting Analysis: A representative lot detected Cas9 in Western Blotting application. (Camargo Ortega, G., et al. (2019). Nature. 567(7746):113-117).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Description de la cible

CRISPR-associated endonuclease Cas9/Csn1 (UniProt: Q99ZW2; also known as SpCas9, SpyCas9, Cas9) is encoded by the Cas9 gene (Gene ID: 69900935) in Streptococcus pyogenes. CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements, and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. Cas9 has a bi-lobed architecture with a recognition lobe (residues 41-425) and a discontinuous nuclease lobe (NUC, residues 1-40 and 453-1053); the crRNA-target DNA lies in a channel between the 2 lobes. The NUC lobe has 2 endonuclease domains. The discontinuous RuvC-like domain in NUC cleaves the target DNA non-complementary to crRNA while the HNH nuclease domain cleaves the target DNA complementary to crRNA. Cas9 serves as a RNA-guided DNA endonuclease that cleaves double-stranded DNA targets with a protospacer adjacent motif (PAM) and complementarity to the guide RNA. (Ref.: Fonfara, I., et al. (2014). Nucleic Acids Res. 42(4); 2577-2590; Jinek, M., et al. (2012). Science. 337(6096); 816-821).

Forme physique

Purified rat monoclonal antibody IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Stockage et stabilité

Recommended storage: +2°C to +8°C.

Autres remarques

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Clause de non-responsabilité

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Code de la classe de stockage

10 - Combustible liquids

Classe de danger pour l'eau (WGK)

WGK 1

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


Certificats d'analyse (COA)

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