PCIF1/CAPAM antibody (pAb)

Application Notes

Applications Validated by Active Motif:
IP: 10 µl per IP
WB*: 0.5-2 µg/mL

*Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot.

For optimal IP results, the addition of 0.05% Tween 20 to all blocking solutions may also reduce background. Individual optimization may be required.

Immunogen

This antibody was raised against a peptide within the C-terminal region of human PCIF1.

Buffer

Purified IgG in PBS, 30% glycerol, and 0.035% sodium azide. Sodium azide is highly toxic.

Background

The abundant modification N6-methyladenosine (m6A), in eukarytic mRNAs, has been identified as a major epitranscriptome mark that plays critical roles in RNA metabolism and function.  In addition to the internal m6A, N6, 2-O dimethyladenosine (m6Am) is present at the transcription start site of capped mRNAs in vertebrates. The abundance of this modification in the transcriptome has been quantified and identified as the writer protein, cap-specific adenosine methyltransferase (CAPAM/PCIF1), is needed for this modification.

CAPAM/PCIF1 is identified as the Methyltransferase responsible for methylating 2-O-methyladenosine (Am) at its N6 position to generate m6Am. m6Am is involved in regulating translation of capped mRNAs. Depleting CAPAM/PCIF1 does not affect mRNA translation but is associated with reduced stability of a subset of m6Am annotated mRNAs.

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

• ChIP = Chromatin Immunoprecipitation;
• ChIP = ChIP Sequencing;
• CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
• CUT&Tag = Cleavage Under Targets and Tagmentation;
• DB = Dot Blot;
• ELISA = Enzyme-linked Immunosorbent Assay;
• EMSA = Electrophoretic Mobility Shift Assay
• FC = Flow Cytometry;
• ICC = Immunocytochemistry;
• IF = Immunofluorescence;
• IHC = Immunohistochemistry;
• IP = Immunoprecipitation;
• MeDIP = Methyl-DNA Immunoprecipitation;
• NEU = Neutralizing;
• WB = Western Blotting