Mouse anti-Human Cas9 Antibody | anti-Cas9 antibody

AbFlex Cas9 antibody (rAb)

Cat. Num. AAA389444

• Reactivity: Human
• Applications: Chromatin Immunoprecipitation, Immunoprecipitation, Chromatin Immunoprecipitation, Immunoprecipitation, ELISA, Western Blot
• Purity: Ni-NTA
• Synonyms: Cas9; AbFlex Cas9 antibody (rAb); CRISPR/Cas9; anti-Cas9 antibody

• Host: Mouse
• Reactivity: Human
• Isotype: IgG2a
• Purity/Purification: Ni-NTA
• Form/Format: Purified IgG in 200mM Hepes pH7.5, 100mM NaCl, 50mM NaOAc, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.
• Concentration: 1ug/ul (varies by lot)

• Applicable Applications for anti-Cas9 antibody: Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation-Seq (ChIP-Seq), ELISA (EIA), Western Blot (WB)
• Application Notes: Applications Validated:; ChIP-Seq: 4ug per ChIP; Bead-based ELISA: 9-75 ng/ml; WB: 2ug/ml; AbFlex recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental clone
• Antibody Type: Recombinant Antibody
• Immunogen: This antibody was raised against a recombinant protein within the N-terminal region of Streptococcus pyogene Cas9. This antibody recognizes Cas9 and dCas9.
• Preparation and Storage: Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20 degree C for up to 2 years. Keep all reagents on ice when not in storage.1ug/ul

SDS-Page

(AbFlex Cas9 antibody (rAb) tested by Western blot. 50 ng recombinant Cas9 protein was probed with 0.5 ug/ml AbFlex Cas9 antibody (Lane 1). HEK293T cells were transfected with a plasmid containing a mammalian expression construct for dCas9 (S. pyogenes). Chromatin was prepared 48 hours post-transfection, boiled and 15 ul (~200,000 cell equivalents) was run on a SDS-PAGE gel and probed with 2 ug/ml AbFlex Cas9 antibody (Lane 2). A molecular weight marker was run in Lane 3.)

Testing Data

(AbFlex Cas9 antibody (rAb) tested by bead-based specificity analysis. Luminex bead-based specificity analysis was used to confirm the specificity of AbFlex Cas9 antibody (rAb) antibody for Cas9. Various proteins were conjugated to MagPlex Luminex beads and incubated with various amounts of AbFlex Cas9 antibody (rAb). Protein-bound antibody was detected with anti-mouse IgG-Phycoerythrin and read in a Luminex instrument. Luminex is a registered trademark of Luminex Corporation.)

Chromatin Immunoprecipitation (ChIP)

(AbFlex Cas9 recombinant antibody (rAb) tested by ChIP-Seq Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT High Sensitivity Kit with 30 ug of Jurkat cell chromatin and 4 ug of AbFlex Cas9 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 8 million sequence tags were mapped to identify binding sites on chromosome 4.)

Related Product Information for anti-Cas9 antibody

Short Description: AbFlex antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. Our AbFlex Cas9 antibody (rAb) was raised in a Mouse host. It has been validated for use in Chromatin Immunoprecipitation, ChIP-Seq, ELISA and Western blot; it has been shown to react with Human samples.

Background: AbFlex antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-tag, a Biotinylation Tag for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex Cas9 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells. Cas9 is a nuclease from Streptococcus pyogenes that can be targeted to particular DNA sequences through a guide RNA that results in double-stranded breaks in DNA. Cas9 is part of the CRISPR/Cas9 gene-editing system that can create a DNA break at a specific location with the genome. 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. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA) Probable. In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently 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, then trimmed by 3'-5' exonucleolytically. DNA-binding requires protein and both RNA species. Cas9 probably recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself.

Product Categories/Family for anti-Cas9 antibody

Primary Antibodies

NCBI and Uniprot Product Information

• Molecular Weight: 160kDa
• Protein Family: CRISPR-associated endonuclease