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AbFlex Cas9 antibody (rAb)

Host / Isotype

Mouse / IgG2a

Reactivity

Human

Applications

ChIP, ChIP-Seq, ELISA, WB

Cat No : 91123,91124 91123

Synonyms

Cas9, Streptococcus pyogenes, CRISPR, clustered regularly interspaced short palindromic repeat, gene-editing, guide RNA, nuclease, endonuclease, targeting, DNA, double-strand break, RNA, genome editing, wb, ip, if, western blotting, immunoprecipitation, immunoflourescence, antibody, antibodies, sample, recombinant, rAb, rec Ab, AbFlex

验证数据展示

  • 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.

    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.

  • 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.

    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.

  • AbFlex Cas9 recombinant antibody (rAb) tested by ChIP-Seq Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) 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.

    AbFlex Cas9 recombinant antibody (rAb) tested by ChIP-Seq Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT High Sensitivity Kit (Cat. No. 53040) 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.

产品信息

Tested Applications ChIP, ChIP-Seq, ELISA, WB

Applications Validated by Active Motif: ChIP-Seq: 4 ug per ChIP Bead-based ELISA: 9 - 75 ng/ml WB: 2 ug/ml AbFlex recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones. For details on the parental clone, see Catalog No. 61577.
Tested Reactivity Human
Host / Isotype Mouse / IgG2a
Class Recombinant
Type 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.
Full Name AbFlex Cas9 antibody (rAb)
Synonyms Cas9, Streptococcus pyogenes, CRISPR, clustered regularly interspaced short palindromic repeat, gene-editing, guide RNA, nuclease, endonuclease, targeting, DNA, double-strand break, RNA, genome editing, wb, ip, if, western blotting, immunoprecipitation, immunoflourescence, antibody, antibodies, sample, recombinant, rAb, rec Ab, AbFlex
Molecular weight 160 kDa
GenBank accession numberWP_014407541
RRIDAB_2793783
Purification Method Ni-NTA
Buffer Purified IgG in 200 mM Hepes pH 7.5, 100 mm NaCl, 50 mM NaOAc, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.
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°C for up to 2 years. Keep all reagents on ice when not in storage.

背景介绍

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.