Cas9 (7A9-3A3) Mouse mAb (Alexa Fluor® 555 Conjugate) #46048
Filter:
- IF
- F
Supporting Data
REACTIVITY | All |
SENSITIVITY | Transfected Only |
MW (kDa) | |
Source/Isotype | Mouse IgG1 |
Application Key:
- IF-Immunofluorescence
- F-Flow Cytometry
Species Cross-Reactivity Key:
- All-All Species Expected
Product Information
Product Description
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 555 fluorescent dye and tested in-house for direct flow cytometric and immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Cas9 (7A9-3A3) Mouse mAb #14697.
Product Usage Information
Application | Dilution |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:50 |
Flow Cytometry (Fixed/Permeabilized) | 1:200 |
Storage
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
Protocol
Specificity / Sensitivity
Cas9 (7A9-3A3) Mouse mAb (Alexa Fluor® 555 Conjugate) recognizes transfected levels of total Cas9 protein.
Species Reactivity:
All Species Expected
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of Cas9 from Streptococcus pyogenes.
Background
The CRISPR associated protein 9 (Cas9) is an RNA-guided DNA nuclease and part of the Streptococcus pyogenes CRISPR antiviral immunity system that provides adaptive immunity against extrachromosomal genetic material (1). The CRISPR antiviral mechanism of action involves three steps: (i), acquisition of foreign DNA by host bacterium; (ii), synthesis and maturation of CRISPR RNA (crRNA) followed by the formation of RNA-Cas nuclease protein complexes; and (iii), target interference through recognition of foreign DNA by the complex and its cleavage by Cas nuclease activity (2). The type II CRISPR/Cas antiviral immunity system provides a powerful tool for precise genome editing and has potential for specific gene regulation and therapeutic applications (3). The Cas9 protein and a guide RNA consisting of a fusion between a crRNA and a trans-activating crRNA (tracrRNA) must be introduced or expressed in a cell. A 20-nucleotide sequence at the 5' end of the guide RNA directs Cas9 to a specific DNA target site. As a result, Cas9 can be "programmed" to cut various DNA sites both in vitro and in cells and organisms. CRISPR/Cas9 genome editing tools have been used in many organisms, including mouse and human cells (4,5). Research studies demonstrate that CRISPR can be used to generate mutant alleles or reporter genes in rodents and primate embryonic stem cells (6-8).
- Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
- Wiedenheft, B. et al. (2012) Nature 482, 331-8.
- Singh, P. et al. (2015) Genetics 199, 1-15.
- Cong, L. et al. (2013) Science 339, 819-23.
- Mali, P. et al. (2013) Science 339, 823-6.
- Li, D. et al. (2013) Nat Biotechnol 31, 681-3.
- Shen, B. et al. (2013) Cell Res 23, 720-3.
- Niu, Y. et al. (2014) Cell 156, 836-43.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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