The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 438 kDa (predicted molecular weight: 438 kDa).
Use 5 µg for 25 µg of chromatin.
TRRAP (TRansformation/tRanscription domain Associated Protein) is an adapter protein that is found in a number of multiprotein chromatin complexes with histone acetyltransferase (HAT) activity. These complexes include PCAF, TFTC-HAT, TIP60 HAT, STAGA, and BAF53 complexes. TRRAP is thought to be responsible for the concerted and context-dependent recruitment of these complexes to chromatin during transcription, replication and DNA repair. TRRAP plays a key role in transcriptional activation by c-Myc, p53/TP53, E2F1 and E2F4.
Transactivation / transformation domain associated protein antibody
Transactivation/transformation domain associated protein antibody
Transformation/transcription domain associated protein antibody
Western blot - TRRAP antibody (ab73546)
All lanes : Anti-TRRAP antibody - ChIP Grade (ab73546) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 2 : Jurkat nuclear extract lysate (ab14844)
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 438 kDa Observed band size : 438 kDa Additional bands at : 150 kDa,90 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 3 minutes
- Anti-TRRAP antibody (ab73546)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab73546 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach for both the active and heterochromatic loci). Primers are located in the first kb of the transcribed region.