Cas9

product

Anti-CRISPR-Cas9 Rabbit Monoclonal Antibody
Boster
catalog: M30929-1

Western blot analysis of CRISPR-Cas9 expression in (1) 293T cell lysate transfected with CRISPR-Cas9; (2) 293T cell lysate; (3) 3T3 cell lysate; (4) PC12 cell lysate (M30929-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-cas9 monoclonal antibody (Catalog # M30929-1) overnight at 4â„ƒ, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for cas9
quantity: 100 ug/vial
price: 299 USD
to the supplier
Cas9 Monoclonal Antibody (7A9)
Invitrogen
catalog: MA5-23519

total number of products: 10

citations: 1

quantity: 50µg
price: US 390
to the supplier
Cas9 Recombinant Rabbit Monoclonal Antibody (JM11-55)
Invitrogen
catalog: MA5-32636

total number of products: 10

quantity: 100µL
price: US 345
to the supplier
Cas9 Polyclonal Antibody
Invitrogen
catalog: PA5-90171

total number of products: 10

quantity: 100µL
price: US 345
to the supplier
CRISPR/Cas9 Antibody [7A9-3A3]
ImmuQuest
catalog: IQ651

quantity: 0.1 ml
price: 345 USD
to the supplier
Anti-CRISPR-Cas9 antibody [EPR19795]
Abcam
catalog: ab203943

total number of products: 19

quantity: 40 µl, 100 µl
price:
to the supplier
Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free
Abcam
catalog: ab218654

total number of products: 19

quantity: 100 µg
price:
to the supplier
Anti-CRISPR-Cas9 antibody [EPR19799] (Phycoerythrin)
Abcam
catalog: ab215701

total number of products: 19

quantity: 100 µl
price:
to the supplier
Cas9 monoclonal antibody (7A9)
Enzo Life Sciences
catalog: ENZ-ABS294-0100

Figure 1. Western blot. HEK293 cells were transiently transfected with pMLM3613myc and lysed in hot Laemmli buffer 40 hours post transfection. Proteins were separated by 7.5% SDS-PAGE and transfered to nitrocellulose. Membranes were blocked in 3% dry milk (NFDM) in PBS-Tween for 1 h at RT and incubated with the indicated antibodies diluted in 0.5% NFDM in PBS-T for 2 hours at RT, followed by incubation with HRP-coupled anti-mouse antibody together with antiBLUE protein marker antibody for 1 hour at RT. Proteins were visualized by ECL.

Figure 2. Immunofluorescence analysis. Hela cells were transiently transfected with an N-terminally Flag-tagged S. pyogenes Cas9 expression vector. 48 hours post transfection cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X and blocked in 2% BSA in PBS for 2 hours at RT. Cells were stained with the indicated antibodies at 4C o/n, followed by incubation with anti mouse-AF488 coupled secondary antibody for 1 h at RT. Nuclei were counter-stained with Hoechst 33342. Images were taken with an Axiovert 135 TV fluorescence microscope. For presentation purposes, images were adjusted for brightness and contrast.

Figure 3. Immunoprecipitation. HEK293T cells expressing N-terminally Flag-tagged S. pyogenes Cas9 were lysed 72 hours post transfection by resuspending the cells in Hunt buffer and subjecting them to three freeze-thaw cycles in liquid nitrogen/on ice. Proteins were immunoprecipitated from 100 ?g of whole cell lysate either for 1 hour at 4°C with crude Cas9 clone 7A9 cell culture supernatant (IgG concentration n.d., lane 3) followed by incubation for 1 hour at 4°C with a 1:1 mixture of protein A/G sepharose beads, or for 2 hours at 4°C with Cas9 clone 7A9 monoclonal antibody crosslinked to a 1:1 mixture of protein A/G sepharose beads. Beads were washed two times with Hunt buffer and once with TBS. Bound proteins were eluted by boiling in Laemmli buffer, separated by 7.5% SDS-PAGE and transfered to nitrocellulose. Membranes were blocked in 3% dry milk (NFDM) in PBS-Tween for 1 h at RT and incubated with Cas9 7A9 monoclonal antibody diluted in 0.5% NFDM in PBS-T for 1 hour at RT, followed by incubation with HRP-coupled anti-mouse antibody. Proteins were visualized by ECL. Asterisk indicates IgG heavy chain.
quantity: 100 µg
price: 365 USD
to the supplier
Streptococcus pyogenes CRISPR-associated endonuclease Cas9/Csn1 Antibody (aa1150 ...
LifeSpan Biosciences
catalog: LS-C340585

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCAS-Guide vector (GE100002, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CAS9.
quantity:
price:
to the supplier
Cas9 (7A9) Antibody
Santa Cruz Biotechnology
catalog: sc-517237

quantity: 100 µg/ml
price: 279 USD
to the supplier
Rabbit polyclonal CAS9 Antibody
OriGene
catalog: TA190309

quantity: 100 µl
price: 345 USD
to the supplier
Anti-CRISPR/Cas9 antibody, Mouse monoclonal
MilliporeSigma
catalog: SAB4200701

citations: 1

quantity:
price:
to the supplier
CRISPR/Cas9 monoclonal antibody
Diagenode
catalog: C15200229

total number of products: 2

citations: 1

quantity:
price:
to the supplier
pX330-U6-Chimeric_BB-CBh-hSpCas9
Addgene
catalog: 42230

total number of products: 69

citations: 800

quantity:
price:
to the supplier

method

CRISPR and Genomic Engineering

other reagents

questions and comments