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MABS1291

Sigma-Aldrich

Anti-Cas9 Antibody, C-term. clone 10C11-A12

clone 10C11-A12, from mouse

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Synonym(s):
CRISPR-associated endonuclease Cas9/Csn1, SpyCas9
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

10C11-A12, monoclonal

species reactivity

bacteria

technique(s)

immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

unmodified

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This Item
MABE1668ABC1490ABS2202
vibrant-m

ABS2202

Anti-Cas9

species reactivity

bacteria

species reactivity

Streptococcus pyogenes

species reactivity

Streptococcus pyogenes

species reactivity

Streptococcus pyogenes, bacteria

antibody form

purified antibody

antibody form

purified immunoglobulin

antibody form

affinity isolated antibody

antibody form

affinity isolated antibody

biological source

mouse

biological source

mouse

biological source

rabbit

biological source

rabbit

UniProt accession no.

Q99ZW2

UniProt accession no.

Q99ZW2

UniProt accession no.

Q99ZW2

UniProt accession no.

Q99ZW2

isotype

IgG1κ

isotype

IgG1κ

isotype

-

isotype

-

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General description

CRISPR-associated endonuclease Cas9/Csn1 (UniProt Q99ZW2; also known as SpyCas9) is encoded by the Streptococcus pyogenes serotype M1 gene cas9 (also known as csn1) gene (Gene ID 901176). Clustered regularly interspaced short palindromic repeats (CRISPRs) are DNA loci containing short repetitions of base sequences complementary to antecedent mobile elements and target invading nucleic acids. Each repetition is followed by short segments of spacer DNA from previous exposures to a virus. Cas9 is an essential endonuclease in Streptococcus pyogenes serotype M1′s CRISPR immune system that confers resistance to foreign genetic elements such as plasmids and phages. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems, correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc), and Cas9. Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, and then trimmed 3′-5′ exonucleolytically.

Specificity

Clone 10C11-A12 recognizes an epitope located in the C-terminal region of Cas9.

Immunogen

His-tagged recombinant S.pyogenes Cas9 C-terminal fragment.

Application

Anti-Cas9 Antibody, C-term. clone 10C11-A12 is an antibody against Cas9 for use in Immunocytochemistry, Immunoprecipitation, and Western Blotting.
Immunocytochemistry Analysis: 33.5 µg/mL from a representative lot detected doxycycline-induced expression of exogenously transfected S. pyogenes Cas9 construct under the control of the PTight (Tet-ON) promoter by fluorescent immunocytochemistry staining of methanol-fixed, acetone-permeabilized HeLa transfectants (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Immunoprecipitation Analysis: 13.4 µg from a representative lot immunoprecipitated exogenously expressed Cas9 in 250 µg of lysate from transfected, but not untransfected, HeLa cells (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Western Blotting Analysis: 1.675 µg/mL from a representative lot detected exogenously expressed Cas9 in 20 µg of lysate from transfected, but not untransfected, HeLa cells (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).
Western Blotting Analysis: Clone 10C11-A12 hybridoma culture supernatant detected doxycycline treatment-induced expression of a FLAG-tagged Cas9 construct under the control of the PTight promoter in a HeLa-derived Tet-ON cell line (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria).

Quality

Evaluated by Western Blotting in Cas9-transfected HeLa cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected the exogenously expressed Cas9 in 10 µg lysate from transfected HeLa cells.

Target description

~158 kDa observed. 158.4 kDa calculated.

Physical form

Format: Purified

Other Notes

Concentration: Please refer to lot specific datasheet.

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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