CRISPR-Cas9 Mouse Monoclonal Antibody [8-A6-6]
Catalog# EM50804
CRISPR-Cas9 Mouse Monoclonal Antibody [8-A6-6]
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WB
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IF-Cell
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Staphylococcus aureus
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unconjugated
Overview
Product Name
CRISPR-Cas9 Mouse Monoclonal Antibody [8-A6-6]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within Streptococcus Pyogenes CRISPR-Cas9 aa1-200.
Species Reactivity
Staphylococcus aureus
Validated Applications
WB, IF-Cell
Positive Control
Human 293T cells transfected with CRISPR-Cas9 lysate.
Conjugation
unconjugated
Clone Number
8-A6-6
RRID
Product Features
Form
Liquid
Concentration
2 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgM
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:200-1:1,000
Target
Function
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Type II system is currently the most commonly used RNA-Guided Endonuclease technology for genome engineering. There are two distinct components to this system: (1) a guide RNA and (2) an endonuclease, in this case the CRISPR associated (Cas) nuclease, Cas9. The gRNA/Cas9 complex is recruited to the target sequence by the base-pairing between the gRNA sequence and the complement to the target sequence in the genomic DNA. For successful binding of Cas9, the genomic target sequence must also contain the correct Protospacer Adjacent Motiff (PAM) sequence immediately following the target sequence.
Background References
1. Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Science. 2013 Dec 12.
2. Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24.
Synonyms
Cas9 antibody
CRISPR-associated endonuclease Cas9/Csn1 antibody
CRISPR-Cas9/Csn1 antibody
csn1 antibody
SpyCas9 antibody
Images
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Western blot analysis of CRISPR-Cas9 on different lysates with Mouse anti-CRISPR-Cas9 antibody (EM50804) at 1/1,000 dilution.
Lane 1: 293T (Human embryonic kidney cell) cell lysate
Lane 2: 293T-OE cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 46 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM50804, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 158 kDa
Observed band size: 158 kDa -
☑ Cell treatment (CT)
Immunofluorescence staining CRISPR-Cas9 in CRISPR-Cas9 transfected 293T cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of 293T cells transfected with or without px459 labeling CRISPR-Cas9 with Mouse anti-CRISPR-Cas9 antibody (EM50804) at 1/1,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-CRISPR-Cas9 antibody (EM50804) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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