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Invitrogen
Staphylococcus aureus Cas9 Monoclonal Antibody (6H4), Alexa Fluor™ 647, eBioscience™
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FIGURE: 1 / 2
Staphylococcus aureus Cas9 Antibody (51-6499-82) in ICC/IF
Immunofluorescent analysis of transgenic SaCas9 expression (cyan) in A549 cells (60% confluency). The cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at 37 C and blocked with 10% Normal Donkey Serum in PBS with 0.1% Triton X-100 for 1 hour at room temperature. Cells were stained with 2.5 µg/mL SaCas9 Mouse Monoclonal Antibody,... View More
Product Details
51-6499-82
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Bacteria
Host/Isotype
Mouse
/ IgG2b, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
6H4
Immunogen
His-tagged recombinant Cas-9 from Staphylococcus aureus (maps to N-terminus)
Conjugate
Excitation/Emission Max
650/671 nm
View spectra
Form
Liquid
Concentration
0.2 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
AB_2815196
Product Specific Information
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.
Target Information
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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