General description
Replication protein A 32 kDa subunit (UniProt: P15927; also known as RP-A p32, Replication factor A protein 2, RPA2, RF-A protein 2, Replication protein A 34 kDa subunit, RP-A p34) is encoded by the RPA2 (also known as REPA2, RPA32, RPA34) gene (Gene ID: 6118) in human. RPA2 is a part of the heterotrimeric replication protein A complex (RPA/RP-A) that binds and stabilizes single-stranded DNA intermediates that form during DNA replication or upon DNA stress. It prevents their reannealing and in parallel, recruits and activates different proteins and complexes involved in DNA metabolism. It also plays an essential role in DNA replication. It is translationally up-regulated in response to DNA damage. In the cellular response to DNA damage, the RPA complex controls DNA repair and DNA damage checkpoint activation. Through recruitment of ATR interacting protein (ATRIP), it activates the ATR kinase. It is also required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin in response to DNA damage. It also recruits XPA and XPG that are involved in nucleotide excision repair. RPA2 is differentially phosphorylated throughout the cell cycle, becoming phosphorylated at the G1-S transition, and dephosphorylated in late mitosis. It is mainly phosphorylated at serine 23 and 29 by cyclin A-CDK2 and cyclin B-CDK1, respectively. It becomes hyperphosphorylated on additional residues including serine 4 and 8, threonine 21, and serine 33 in response to DNA damage. This hyperphosphorylation is mediated by ATM, ATR, DNA-PK. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Yin, J-Y., et al. (2013). Carcinogenesis. 34(6); 1224-1231; Sleeth, KM., et al. (2007). J. Mol. Biol. 373(1); 38-47; Erdile, LF., et al. (1990). J. Biol. Chem. 265(6); 3177-3182).
We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.
ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Immunogen
KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphoserines 4 and 8 from the N-terminal region of human Replication protein A 32 kDa subunit (RPA2).
Application
Quality Control Testing
Evaluated by Western Blotting in lysate from HeLa cells treated with Etoposide.
Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected RPA2 phosphorylated on serine 4 and 8 in lysate from HeLa cells, overnight serum starved and treated with Etoposide ( 20 g/mL; 3 h)
Tested applications
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Phospho-RPA2-Ser4/8 in human lung cancer tissue sections.
Affinity Binding Assay: A representative lot of this antibody bound Phospho-RPA32-Ser4/8 peptide with a KD of 2.7 x 10-7 in an affinity binding assay.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Evaluated by Western Blotting in lysate from HeLa cells treated with Etoposide.
Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected RPA2 phosphorylated on serine 4 and 8 in lysate from HeLa cells, overnight serum starved and treated with Etoposide ( 20 g/mL; 3 h).
Biochem/physiol Actions
Clone 2F23 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Replication protein A 32 kDa subunit (RPA2) phosphorylated on serine 4 and 8.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.
Preparation Note
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Segment | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 2F23, recombinant monoclonal |
| description | recombinant, expressed in HEK 293 cells |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt 29.25 kDa,observed mol wt ~35 and 38 kDa |
| purified by | using Protein A |
| species reactivity | human |
| packaging | antibody small pack of 25 μL |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| sustainability | Greener Alternative Product |
| technique(s) | affinity binding assay: suitable,immunohistochemistry: suitable,western blot: suitable |
| isotype | IgG |
| epitope sequence | N-terminal |
| Protein ID accession no. | NP_002937 |
| UniProt accession no. | P15927 |
| greener alternative category | , Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
| target post-translational modification | phosphorylation (pSer4/8) |
| Gene Information | human ... RPA2(6118) |
