L1/ORF2 antibody

Antigen: L1/ORF2

Clonality: Polyclonal

Host: Chicken

Reactivity: Human, Chimpanzee

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP), Immunofluorescence (IF)

Catalog no.: ABIN349625

Additional Information

Gene ID: AAD39215

Immunogen: This IgY fraction antibody was prepared from eggs of chickens laid after repeated immunizations with two synthetic peptides conjugated to keyhole limpet hemocyanin (KLH). The peptides correspond to regions within the endonuclease domain of L1/ORF2 protein.

Format: Liquid (sterile filtered)

Description: The L1 (LINE-1), or Long INterspersed Element , retrotransposon ORF2 is the most common open reading frame in the human genome, present in various forms in many thousands of copies. This large family of proteins includes magnesium dependent endonucleases and a large number of phosphatases involved in intracellular signaling. Both intact ORF1 and ORF2 are absolutely required for autonomous retro-transposition. ORF2 encodes an endonuclease, reverse transcriptase, and zinc knuckle domains. The expression of ORF2 appears to be tightly regulated except in germ line tissues, embryonic tissues, and certain cancers including teratomas, testicular cancers, and leukemias. This antibody is intended for use in studying control of L1 retrotransposons.

Specificity: This IgY fraction antibody is directed against two regions within the endonuclease domain of L1 ORF2 protein. This product is an IgY fraction antibody purified from monospecific chicken egg yolks by a multi-step process which includes selective precipitation and salt fractionation followed by extensive dialysis against the buffer stated above. Reactivity occurs against human L1/ORF2 protein and is useful in determining its presence in various assays. A BLAST analysis was used to suggest cross reactivity with L1/ORF2 proteins from chimpanzee sources based on 100% homology with the immunizing sequences, and from macaque, fruit fly, cattle, dog, opossum, and rat sources based on 69 - 88% homology with the immunizing sequences.

Application Details

Application Notes: Recommended Dilutions: User optimized

Concentration: 1.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20°C or below prior to opening. Dilute only prior to immediate use. For extended storage, aliquot contents and freeze at -20°C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of opening.

Restrictions: For Research Use only

Images

Western blot using IgY fraction of anti-L1/ORF2 antibody shows detection of induced bacterially expressed human ORF2 (left lane). No specific band staining is seen in the uninduced lane (right lane). The lower molecular weight bands represent non-specific staining. The band at ~70 kDa corresponds to a human L1/ORF2 EN domain fusion protein (arrowhead). Personal communication, D. Symer, NCI, Bethesda, MD.