Detection of RUNX2/CBFA1-regulated Genes by Chromatin Immunoprecipitation.
Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. RUNX2/CBFA1/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human RUNX2/CBFA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2006) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The VEGF promoter was detected by standard PCR.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Core Binding Factor A1 (CBFA1; also called AML-3, PEBP-2 alpha A, OSF-2, RUNX2) is a 56 kDa DNA-binding protein that belongs to the Runt-related family of transcription factors. It is synthesized as a 521 amino acid (aa) protein with multiple domains. From the N-terminus, there is a 19 aa activation domain (AD1), a 41 aa glutamine/alanine (Q/A) repeat segment (AD2), a 128 aa Runt/DNA-binding region, a 9 aa NLS and a 285 aa ProSerThr (PST)-rich sequence that is needed for gene activation. Two isoforms exist, one with a truncated AD1 domain and another with a 22 aa deletion in the AD3 section of PST-rich sequence. Human CBFA1 is 98% aa identical to mouse CBFA1.
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