Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

RPA70 Antibody #2267

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC F H Mk Endogenous 70 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

RPA70 antibody detects endogenous levels of total RPA70 subunit.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids near the amino terminus of human RPA70. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 and K562 cells, using RPA70 Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, using RPA70 antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent images of HeLa cells, untreated (left) or UV-treated (right), labeled with RPA70 Antibody (green) showing translocation to distinct nuclear foci after UV damage. Actin filaments have been labeled with Alexa Fluor® 555 phalloidin. Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


Background

RPA70 (HSSB, REPA1, RF-A, RP-A, p70) is a component of a heterotrimeric complex, composed of 70, 32/30 and 14 kDa subunits, collectively known as RPA. RPA is a single stranded DNA binding protein, whose DNA binding activity is believed to reside entirely in the 70 kDa subunit. The complex is required for almost all aspects of cellular DNA metabolism such as DNA replication (1-3), recombination, cell cycle and DNA damage checkpoints, and all major types of DNA repair including nucleotide excision, base excision, mismatch and double-strand break repairs (4-7). In response to genotoxic stress in eukaryotic cells, RPA has been shown to associate with the Rad9/Rad1/Hus1 (9-1-1) checkpoint complex (8). RPA is hyperphosphorylated upon DNA damage or replication stress by checkpoint kinases including ataxia telangiectasia mutated (ATM), ATM and Rad3-related (ATR), and DNA-dependent protein kinase (DNA-PK) (9-11). Hyperphosphorylation may alter RPA-DNA and RPA-protein interactions. In addition to the checkpoint partners, RPA interacts with a wide variety of protein partners, including proteins required for normal replication such as RCF, PCNA and Pol α, and also proteins involved in SV40 replication, such as DNA polymerase I and SV40 large T antigen (10,12).

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  6. Kastan, M.B. and Bartek, J. (2004) Nature 432, 316-323.
  7. Sancar, A. et al. (2004) Annu. Rev. Biochem. 73, 39-85.
  8. Guo, S. et al. (2006) J Biol Chem 281, 21607-16.
  9. Wu, X. et al. (2005) Oncogene 24, 4728-4735.
  10. Binz, S.K. et al. DNA Repair (Amst) 3, 1015-1024.
  11. Nuss, J.E. et al. (2005) Biochemistry 44, 8428-8437.
  12. Yuzhakov, A. et al. (1999) EMBO J. 18, 6189-6199.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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