The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50000 - 1/200000. Detects a band of approximately 32 kDa (predicted molecular weight: 29 kDa).
The antibody has been tested with milk (5%) and BSA (2%) as blocking reagents, but it provided better results with 5%milk.
1/10 - 1/100.
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
Required for DNA recombination, repair and replication. The activity of RP-A is mediated by single-stranded DNA binding and protein interactions. Functions as component of the alternative replication protein A complex (aRPA). aRPA binds single-stranded DNA and probably plays a role in DNA repair; it does not support chromosomal DNA replication and cell cycle progression through S-phase. In vitro, aRPA cannot promote efficient priming by DNA polymerase alpha but supports DNA polymerase delta synthesis in the presence of PCNA and replication factor C (RFC), the dual incision/excision reaction of nucleotide excision repair and RAD51-dependent strand exchange.
Phosphorylated in a cell-cycle-dependent manner (from the S phase until mitosis). Phosphorylated by ATR upon DNA damage, which promotes its translocation to nuclear foci. Can be phosphorylated in vitro by PRKDC/DNA-PK in the presence of Ku and DNA, and by CDK1.
Nucleus. Nucleus > PML body. Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage.
Dot Blot - Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
Dot blot analysis of RPA32/RPA2 (pT21) phospho peptide (lane 1) and RPA32/RPA2 non-phospho peptide (lane 2) labelling RPA32/RPA2 (phospho T21) with ab109394 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Western blot - RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/50000 dilution
Lane 1 : HeLa cell lysates, untreated Lane 2 : HeLa cell lysates, treated with Camptothecin
Biehs R et al. DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination. Mol Cell65:671-684.e5 (2017).
Read more (PubMed: 28132842) »
Yamamoto K et al. Kinase-dead ATM protein is highly oncogenic and can be preferentially targeted by Topo-isomerase I inhibitors. Elife5:N/A (2016).
Read more (PubMed: 27304073) »