General description
LINE-1 retrotransposable element ORF2 protein (UniProt: O00370; also known as EC:2.7.7.49, ORF2p endonuclease, ORF2p) is encoded by the Orf2p gene (Gene ID: 107987054) in human. Full length long interspersed element-1 (L1) is an autonomous non-long terminal repeat retrotransposon that contains two open reading frames encoding ORF1p and ORF2p proteins. L1 is reported to be responsible for all of the retrotransposon-induced genomic instability in the human genome and is the only active source of the functional ORF1 and ORF2 proteins required for mobilization of LINEs, SINEs, and SVA elements. The human ORF2p, a 149 kDa protein, contains three domains: an N-terminal endonuclease domain, a reverse transcriptase domain, and C-terminal domain with putative RNA binding activity. Its endonuclease activity allows the introduction of nicks in the chromosomal target DNA and is known to cleave DNA in AT-rich regions between a 5′ stretch of purines and a 3′ stretch of pyrimidines, corresponding to sites of LINE-1 integration in the genome. Human and mouse ORF2p is reported to exhibit a high degree of sequence homology. Clone 1G4E11 can recognize sequences within the endonuclease domain of the human ORF2 protein. It exhibits specificity to the human ORF2p and can detect the full-length and truncated ORF2 proteins expressed in mammalian cells. (Ref.: Sokolowski M., et al. (2014). Mob DNA. 5(1); 29; Feng, Q., et al. (1996). Cell. 87(5); 905-916).
Immunogen
A His-tagged recombinant human protein containing an ORF2p endonuclease.
Application
Quality Control Testing
Evaluated by Western Blotting in lysate from HEK293 cells overexpressing human ORF2p EN.
Western Blotting Analysis: A 1:500 dilution of this antibody detected LINE1 ORF2 in lysate from HEK293 cells overexpressing human ORF2p EN.
Tested applications
Western Blotting Analysis: A representative lot detected LINE1 ORF2 in Western Blotting applications (Sokolowski, M. et al. (2014). Mob DNA. 5(1):29.).
Inhibition Analysis: A representative lot partially inhibited L1 endonuclease activity in an in vitro endonuclease cleavage assay. (Sokolowski, M. et al. (2014). Mob DNA. 5(1):29.).
ELISA Analysis: A representative lot detected LINE1 ORF2 in ELISA applications (Sokolowski, M. et al. (2014). Mob DNA. 5(1):29.).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biochem/physiol Actions
Clone 1G4E11 is a mouse monoclonal antibody that detects LINE-1 retrotransposable element ORF2 protein (LINE1 ORF2).
Physical form
Purified mouse monoclonal antibody IgG1 in PBS without azide.
Preparation Note
0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | mouse |
| Quality Segment | 200 |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 1G4E11, monoclonal |
| mol wt | calculated mol wt 149.01 kDa |
| purified by | using protein G |
| species reactivity | human |
| packaging | antibody small pack of 100 μL |
| technique(s) | ELISA: suitable,inhibition assay: suitable,western blot: suitable |
| isotype | IgG1 |
| epitope sequence | N-terminal |
| UniProt accession no. | O00370 |
| storage temp. | -10 to -25°C |
| target post-translational modification | unmodified |
| Gene Information | human ... Orf2p(0) |
