Anti-AsCas12a (Cpf1) antibody, Mouse monoclonal
SIGMA/SAB4200756 - clone AsCpf-11, purified from hybridoma cell culture
Synonym: Anti-AsCpf1 antibody, Mouse monoclonal; Anti-
Product Type: Chemical
Immunofluorescence Human HEK-293T cells over-expressing AsCpf1-GFP were fixed and permeabilized with methanol followed by aceton. Fixed cells were stained with 1 μg/mL Anti-AsCpf1 antibody, Mouse monoclonal, Clone: AsCpf-11 (Cat. No. SAB4200756). The antibody was developed using Goat Anti-Mouse IgG, Cy3™ conjugate (A). Cells were visualized with GFP filter (B) and counterstained with DAPI (blue) to stain nuclei (C). (D) represents merged image.
Western Blotting Recombinant AsCpf1 expressed in E. coli was separated on SDS-PAGE and probed with Anti-AsCpf1 antibody, Mouse monoclonal, Clone: AsCpf-11 (Cat. No. SAB4200756). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. 5 μg/mL antibody 2. 2.5 μg/mL antibody 3. 1.25 μg/mL antibody 4. 0.625 μg/mL antibody 5. Negative control: without first antibody
Western Blotting Recombinant CRISPR proteins expressed in E. coli were separated on SDS-PAGE (0.5 μg/lane) and probed with 1μg/mL Anti-AsCpf1 antibody, Mouse monoclonal, Clone: AsCpf-11 (Cat. No. SAB4200756). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. SpCas9 protein 2. Cas9 nickase protein 3. Dead Cas9 (dCas9) protein 4. FnCas9 protein 5. AsCpf1 protein 6. LbCpf1 protein
Immunoprecipitation Anti-AsCpf1 antibody, Mouse monoclonal, Clone: AsCpf-11 (Cat. No. SAB4200756) was used to immunoprecipitate AsCpf1 from whole extract of Human HEK-293Tcells over-expressing AsCpf1. Detection antibody: Anti-AsCpf1 antibody, Mouse monoclonal, Clone AsCpf-11 (Cat. No. SAB4200756), followed by Goat-Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. 2.5 μg/test antibody 2. 5 μg/test antibody 3. Negative control: 5 μg/test without lysate 4. Negative control: 5 μg/test non-relevant antibody 5. Negative control: without antibody 6. Whole extract of Human HEK-293T over-expressing AsCpf1
| antibody form | purified from hybridoma cell culture |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | AsCpf-11, monoclonal |
| concentration | ~1.0 mg/mL |
| form | buffered aqueous solution |
| isotype | IgG2a |
| mol wt | ~135 kDa |
| Quality Level | 200  |
| shipped in | dry ice |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunoblotting: 1.25-2.5 μg/mL using purified recombinant AsCpf1 produced in E. coli |
| immunofluorescence: 1.25-2.5 μg/mL using human HEK-293T cells over-expressing AsCpf1 protein | |
| immunoprecipitation (IP): 2.5-5 μg/test using lysate of human HEK-293T cells over-expressing AsCpf1 protein | |
| UniProt accession no. | U2UMQ6  |
| Application: | Monoclonal Anti-AsCpf1 recognizes Cpf1 from Acidaminococcus sp. (strain BV3L6). The product has been used in several immunochemical techniques including • immunoblotting (~135 kDa), • immunofluorescence • immunoprecipitation Monoclonal anti-AsCpf1 antibody can provide a useful tool for genome editing research such as detecting and monitoring AsCpf1 positively transfected cells. |
| General description: | Clustered, regularly interspaced, short palindromic repeat (CRISPR) systems, encode RNA-guided endonucleases that are essential for bacterial adaptive immunity. Ddepending on the architecture of the effector-CRISPR RNA (crRNA) interference module, the ddifferent CRISPR-Cas systems could be assigned into two classes: Class-1 systems of multi-subunit complex such as Cascade and Class-2 systems of single enzyme, such as Cas9. Cpf1 (CRISPR from Prevotella and Francisella 1) belongs to Class-2 type V CRISPR-Cas endonuclease system.4-5 Cpf1 comprise several differences from Cas9 protein including cleavage with 5′overhangs, a shorter guide RNA and a longer distance between the seed sequence and the cleavage site. AsCpf1, Cpf1 from Acidaminococcus sp. (strain BV3L6), was examined together with 15 members of Cpf1 nuclease family and shown to mediate efficient genome editing in HEK293FT cells with an improved results compared to SpCas9.5 AsCpf1 crystal structure in a complex with crRNA and partially duplexed target DNA, shows that AsCpf1 acts as a monomer thus identifying a unique mechanism employed by AsCpf1 for target recognition. |
| Immunogen: | recombinant Cpf1 from Acidaminococcus sp. (strain BV3L6) |
| Other Notes: | This product is for R&D use only, not for drug, household, or other uses. In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test. |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide. |
| Storage and Stability: | For continuous use, store at 2-8°C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |