Staphylococcus aureus Cas9 Monoclonal Antibody (6H4), Alexa Fluor™ 647, eBioscience™, Invitrogen™

Description

• Description: This 6H4 monoclonal antibody recognizes Staphylococcus aureus Cas9 endonuclease
• Applications Reported: This 6H4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis
• Applications Tested: This 6H4 antibody has been tested by intracellular staining followed by flow cytometric analysis of Staphylococcus aureus derived Cas9-GFP transfected 293T cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol
• Please refer to Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins located at www.thermofisher.com/flowprotocols
• This may be used at less than or equal to 0.06 μg per test
• A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest
• Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser Cas9 is an RNA-guided DNA endonuclease enzyme associated with the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) adaptive immunity system in bacteria and genetic engineering
• The system is unique and flexible due to its dependence on RNA as the element that targets the nuclease to a desired DNA sequence
• It can be used to induce indel mutations, specific sequence replacements/insertions, and large deletions or genomic rearrangements at any desired location in the genome
• In addition, Cas9 can also be used to mediate upregulation of specific endogenous genes or to alter histone modifications or DNA methylation
• Cas9 may be derived from Streptococcus pyogenes (SpCas9) or Staphylococcus aureus (SaCas9)
• SaCas9 has a number of properties that make it advantageous for genome editing, including its small size, high efficiency, nickase activity, and apparent specificity
• SaCas9 principally recognizes an NNGRRT protospacer adjacent motif (PAM) and cleaves target DNA at rates comparable to SpCas9 with an ideal spacer lengths of 24 to 20 nucleotides
• In comparison to SpCas9, SaCas9 is smaller (1368 AA versus 1053 AA) and generates indels at a comparable rate when directed against target DNA with the mutually permissive NGGRRT PAM
• This makes it the preferred enzyme for all-in-one delivery of Cas9 and multiple gRNA expression cassettes with Adeno-associated vectors.