Anti-LbCas12a (Cpf1) antibody, Mouse monoclonal
SIGMA/SAB4200777 - clone LbCpf1, purified from hybridoma cell culture
Synonym: Anti-
Product Type: Chemical
Immunofluorescence Human HEK-293Tcells over-expressing LbCpf1-GFP were fixed and permeabilized with methanol followed by aceton. Fixed cells were stained with 0.5 μg/mL Anti-LbCpf1 antibody, Mouse monoclonal, Clone: LbCpf1 (Cat. No. SAB4200777). 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 Whole extract of Human HEK-293T cells over-expressing LbCpf1-FLAG was separated on SDS-PAGE and probed with Anti-LbCpf1 antibody, Mouse monoclonal, Clone: LbCpf1 (Cat. No. SAB4200777). The antibody was developed using Goat Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. 1.25 μg/mL antibody 2. 0.625 μg/mL antibody 3. 0.31 μg/mL antibody 4. Positive control: 1:5000 Monoclonal anti-FLAG antibody (Cat. No. F1804) 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 1 μg/mL Anti-LbCpf1 antibody, Mouse monoclonal, Clone: LbCpf1 (Cat. No. SAB4200777). 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-LbCpf1 antibody, Mouse monoclonal, Clone: LbCpf1 (Cat. No. SAB4200777) was used to immunoprecipitate LbCpf1 from whole extract of Human HEK-293T cells over-expressing LbCpf1. Detection antibody: Anti-LbCpf1 antibody, Mouse monoclonal, Clone LbCpf1 (Cat. No. SAB4200777), followed by Goat-Anti-Mouse IgG-Peroxidase (Cat. No. A2304) and a chemiluminescent substrate. Lanes 1. 1 μg/test antibody 2. 2.5 μg/test antibody 3. 5 μg/test antibody 4. Negative control: 5 μg/test without lysate 5. Negative control: 5 μg/test non-relevant antibody 6. Negative control: without antibody 7. Whole extract of Human HEK-293T over-expressing LbCpf1
| antibody form | purified from hybridoma cell culture |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | LbCpf1, monoclonal |
| concentration | ~1.0 mg/mL |
| form | buffered aqueous solution |
| isotype | IgG1 |
| mol wt | ~135 kDa |
| Quality Level | 200  |
| shipped in | dry ice |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunoblotting: 0.6-1.2 μg/mL using human HEK-293T cells over-expressing LbCpf1 protein |
| immunofluorescence: 0.25-0.5 μg/mL using human HEK-293T cells over-expressing LbCpf1 protein | |
| immunoprecipitation (IP): 1-2.5 μg/test using lysate of human HEK-293T cells over-expressing LbCpf1 protein | |
| UniProt accession no. | A0A182DWE3  |
| Application: | Monoclonal Anti-LbCpf1 specifically recognizes Cpf1 from Lachnospiraceae bacterium ND2006. The product may be used in several immunochemical techniques including Immunoblotting (∼135 kDa), Immunofluorescence and Immunoprecipitation. Monoclonal Anti-LbCpf1 does not cross react with Cpf1 from Acidaminococcus sp. (strain BV3L6) ), SpCas9 from Streptococcus pyogenes bacteria and FnCas9 from Francisella novicida bacteria. |
| General description: | Clustered, regularly interspaced, short palindromic repeat (CRISPR) systems encode RNA-guided endonucleases that are essential for bacterial adaptive immunity. Depending on the architecture of the effector-CRISPR RNA (crRNA) interference module, different CRISPR-Cas systems could be assigned into two classes1: 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. 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 cleavage site. LbCpf1, Cpf1 from Lachnospiraceae bacterium ND2006, was examined together with 15 members of Cpf1 nuclease family and proved to mediate efficient genome editing in HEK293FT cells with improved results compared to SpCas9.5 According to thr crystal structure, LbCpf1 has a triangle-shaped architecture with a large positively charged channel at the centre.7 The crRNA binding was shown to induce the pronounced structural rearrangements of LbCpf1, leading to formation of a substrate-binding conformation of LbCpf1. Over-expressed in plant cells, LbCpf1 demonstrated more than 10-fold transcriptional repression of the target gene. Monoclonal anti-LbCpf1 antibody can provide a useful tool for genome editing research, such as detecting and monitoring LbCpf1 positively transfected cells. |
| Immunogen: | Recombinant LbCpf1 |
| 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 |