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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, ICC, IHC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE285 |
| Replaces | NA19L |
| Description | Anti-Replication Protein A Antibody, clone RPA34-20 |
| Alternate Names |
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| Background Information | RPA (RP-A p32) is a heterotrimeric protein complex that binds specifically to single-stranded DNA (ssDNA). It is composed of three subunits: RPA1 (70 kDa), RPA2 (32 kDa), and RPA3 (14 kDa). RPA plays multiple roles in DNA replication. At the onset of DNA replication, RPA is loaded onto chromatin, and is needed for subsequent loading of DNA polymerase and other replication proteins to initiate DNA replication. After replication begins, RPA moves with replication forks, stabilizing ssDNA and assisting in DNA synthesis. In addition to its replication function, RPA is also known to play essential roles in damage repair and recombination. The 32 kDa subunit is phosphorylated by the cdc2 family of kinases when cells enter S-phase; and by ATM, ATR, and DNA-PK proteins in response to DNA damage. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Use Anti-Replication Protein A Antibody, clone RPA34-20 (mouse monoclonal antibody) validated in WB, ICC, IHC to detect Replication Protein A also known as Replication protein A 32 kDa subunit, RP-A p32. |
| Key Applications |
|
| Application Notes | Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Replication Protein A in HeLa and A431 cells. Immunohistochemistry Analysis: A 1:5 dilution from a representative lot detected Replication Protein A in human colorectal adenocarcinoma tissues. |
| Biological Information | |
|---|---|
| Immunogen | Replication Protein A purified from U293 cells |
| Clone | RPA34-20 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Isotype | IgG1κ |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: Required for DNA recombination, repair and replication. The activity of RP-A is mediated by single-stranded DNA binding and protein interactions. Required for the efficient recruitment of the DNA double-strand break repair factor RAD51 to chromatin in response to DNA damage. 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. SUBUNIT STRUCTURE: Heterotrimer of 70, 32 and 14 kDa chains (canonical replication protein A complex). Component of the alternative replication protein A complex (aRPA) composed of RPA1, RPA3 and RPA4. The DNA-binding activity may reside exclusively on the 70 kDa subunit. Binds to SERTAD3/RBT1. Interacts with TIPIN. Directly interacts with PPP4R2, but not with SMEK2; this interaction is DNA damage-dependent and leads RPA2 dephosphorylation by PPP4C recruitment. Interacts with RAD51, preferentially when hyperphosphorylated. Directly interacts with RFWD3. SUBCELLULAR LOCATION: Nucleus. Nucleus > PML body. Note: Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage. POST-TRANSLATIONAL MODIFICATION: Phosphorylated in a cell-cycle-dependent manner (from the S phase until mitosis). In response to DNA damage, recruited to DNA-repair nuclear foci, as an hypophosphorylated form. The necessary dephosphorylation step is catalyzed by PP4. Subsequent hyperphosphorylation, catalyzed by ATR, is required for RAD51 recruitment to chromatin and efficient DNA repair. Can be phosphorylated in vitro by PRKDC/DNA-PK in the presence of Ku and DNA, and by CDK1. Phosphorylation at Thr-21 depends upon RFWD3 presence. |
| Molecular Weight | ~30 kDa observed. Uniprot describes three isoforms at ~29 kDa, ~30 kDa, and ~39 kDa. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blot in HeLa cell lysate. Western Blot Analysis: 1 µg/mL of this antibody detected Replication Protein A in 10 µg of HeLa cell lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |