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Cas9 | CRISPR-associated endonuclease 9 (polyclonal)

265 €
Buy 2 items of this product for 198 €/each
Buy 3 items of this product for 180 €/each

AS16 3690  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Cas9 from Streptococcus thermophilus

PRODUCT INFORMATION IN PDF

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AS16 3690

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product information
Background Cas9 (CRISPR-associated endonuclease 9) is an RNA-guided DNA endonuclease associated with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) type II adaptive immunity system. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). Cas9 protein serves as a genome engineering tool to induce site-directed double strand breaks in DNA.
Immunogen

KLH-conjugated synthetic peptide derived from Streptococcus thermophilus Cas9/Csn1 protein sequence, UniProt: Q03JI6

Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl of sterile water.
Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications

Western Blot (WB)

Related products AS16 3841 | Anti/Cpf1 | CRISPR from Prevotella and Francisella 1, rabbit antibodies

Collection of antibodies to tag proteins

Plant and algal protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution

1 : 1000 (WB)

Expected | apparent MW

Depends upon a MW of a protein which is a fusion partner.

Confirmed reactivity
Predicted reactivity
Not reactive in
Additional information
Selected references

To be added when available, antibody released in March 2017.


Application example


Western blot using polyclonal anti-Cas9 antibodies


Nicotiana benthamiana (N b.): sample from 5 weeks old plants second leaves (previously infiltrated with A.tumefaciens containing Cas9 transformation casettes(Nb+). Hordeum vulgare (Hv 1-4 , Hv- samples): 2 months old plants’ youngest leaves (stably transformed (Hv 1, -2, -3, -4) and wild type (Hv-) plants). 100 mg plant tissue (homogenised in liquid N2) +400 μl extraction buffer (15 min 4 °C 1500rpm centrifugation). Tissue was extracted with following extaction buffer: 10 mM Tris-HCl, 1 mM EDTA, 150 mM NaCl, 10 V% glicerol, 0,5 V% Nonidet P-40, 5 mM NaF. Right before use: 1 mM DTT, 100 mM PMSF, 200 mM Na3VO4 is added to the extraction buffer. Denaturation: sample in extraction buffer + equal volume 2X Laemmli buffer, 72°C, 10 min.
20 μl of the samples were separated on 8% SDS-PAGE, and blotted with transblot turblot turbo (BioRad) to 0.45 um pore size PVDF (Hybond, GE). Blots were blocked with 5% milk powder in PBST for 60 min at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for 160 min at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 2 times for 10 min in PBST buffer at RT with agitation. Blot was incubated in secondary antibody (Goat anti-rabbit from Agrisera, AS09 602) diluted to 1:20 000 in PBST for 90 min at RT with agitation. The blot was washed as above and developed for 1 min with Bio-Rad Clarity Western ECL substrate using Bio-Rad ChemiDocTM Imaging System. Exposure time was 170.6 sec.

Courtesy Dr. Éva Hamar, ABC, Hungary

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