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mouse anti-RPA32 monoclonal antibody (12F3.3) 6624

$503.00

Antibody summary

  • Mouse monoclonal to RPA32
  • Suitable for: WB,ICC/IF,IP
  • Isotype: IgG2a
  • 100 µg
SKU: 6624parent Category: Tag:
Weight1 lbs
Dimensions9 × 5 × 2 in
host

mouse

isotype

IgG2a

clonality

monoclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

RPA32

available sizes

100 µg

mouse anti-RPA32 monoclonal antibody (12F3.3) 6624

antibody
Tested applications
WB,ICC/IF,IP
Recommended dilutions
Immunoblotting,

Immunoprecipitation: use at 0.2-1 ug/mL.

Positive control: HeLa, Raji, and 3T3 cells and recombinant protein.
Immunogen
N-terminally 6x-His tagged fusion protein corresponding to full-length human RPA32 expressed in E. coli.
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -70°C. Avoid multiple freeze-thaw cycles.
Storage buffer
PBS, pH 7.4
Purity
protein affinty purification
Clonality
monoclonal
Isotype
IgG2a
Compatible secondaries
goat anti-mouse IgG, H&L chain specific, peroxidase conjugated polyclonal antibody 5486
goat anti-mouse IgG, H&L chain specific, biotin conjugated, Conjugate polyclonal antibody 2685
goat anti-mouse IgG, H&L chain specific, FITC conjugated polyclonal antibody 7854
goat anti-mouse IgG, H&L chain specific, peroxidase conjugated polyclonal antibody, crossabsorbed 1706
goat anti-mouse IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1716
goat anti-mouse IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1721
Isotype control
Mouse monocolonal IgG2a - Isotype Control
target relevance
Protein names
Replication protein A 32 kDa subunit (RP-A p32) (Replication factor A protein 2) (RF-A protein 2) (Replication protein A 34 kDa subunit) (RP-A p34)
Gene names
RPA2,RPA2 REPA2 RPA32 RPA34
Protein family
Replication factor A protein 2 family
Mass
29247Da
Function
As part of the heterotrimeric replication protein A complex (RPA/RP-A), 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. Thereby, it plays an essential role both in DNA replication and the cellular 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 ATRIP activates the ATR kinase a master regulator of the DNA damage response. It is required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin in response to DNA damage. Also recruits to sites of DNA damage proteins like XPA and XPG that are involved in nucleotide excision repair and is required for this mechanism of DNA repair. Also plays a role in base excision repair (BER) probably through interaction with UNG. Also recruits SMARCAL1/HARP, which is involved in replication fork restart, to sites of DNA damage. May also play a role in telomere maintenance.
Subellular location
Nucleus. Nucleus, PML body. Note=Redistributes to discrete nuclear foci upon DNA damage in an ATR-dependent manner.
Structure
Component of the replication protein A complex (RPA/RP-A), a heterotrimeric complex composed of RPA1, RPA2 and RPA3 (PubMed:2406247, PubMed:19116208, PubMed:10449415). Interacts with PRPF19; the PRP19-CDC5L complex is recruited to the sites of DNA repair where it ubiquitinates the replication protein A complex (RPA) (PubMed:24332808). Interacts with SERTAD3 (PubMed:10982866). Interacts with TIPIN (PubMed:17141802, PubMed:17296725). Interacts with TIMELESS (PubMed:17141802). Interacts with PPP4R2; the interaction is direct, DNA damage-dependent and mediates the recruitment of the PP4 catalytic subunit PPP4C (PubMed:20154705). Interacts (hyperphosphorylated) with RAD51 (PubMed:20154705). Interacts with SMARCAL1; the interaction is direct and mediates the recruitment to the RPA complex of SMARCAL1 (PubMed:19793861, PubMed:19793862, PubMed:19793863). Interacts with RAD52 and XPA; those interactions are direct and associate RAD52 and XPA to the RPA complex (PubMed:7700386, PubMed:8702565, PubMed:17765923, PubMed:11081631). Interacts with FBH1 (PubMed:23319600). Interacts with ETAA1; the interaction is direct and promotes ETAA1 recruitment at stalled replication forks (PubMed:27601467, PubMed:27723720, PubMed:27723717). Interacts with RFWD3 (PubMed:21504906, PubMed:21558276, PubMed:26474068, PubMed:28575657). Interacts with DDI2 (PubMed:29290612). Interacts (in unphosphorylated form via N-terminus) with EIF4EBP3; the interaction enhances EIF4EBP3-mediated inhibition of EIF4E-mediated mRNA nuclear export (PubMed:22684010).
Post-translational modification
Differentially phosphorylated throughout the cell cycle, becoming phosphorylated at the G1-S transition and dephosphorylated in late mitosis. Mainly phosphorylated at Ser-23 and Ser-29, by cyclin A-CDK2 and cyclin B-CDK1, respectively during DNA replication and mitosis. Dephosphorylation may require the serine/threonine-protein phosphatase 4. Phosphorylation at Ser-23 and Ser-29 is a prerequisite for further phosphorylation. Becomes hyperphosphorylated on additional residues including Ser-4, Ser-8, Thr-21 and Ser-33 in response to DNA damage. Hyperphosphorylation is mediated by ATM, ATR and PRKDC. Primarily recruited to DNA repair nuclear foci as a hypophosphorylated form it undergoes subsequent hyperphosphorylation, catalyzed by ATR. Hyperphosphorylation is required for RAD51 recruitment to chromatin and efficient DNA repair. Phosphorylation at Thr-21 depends upon RFWD3 presence.; DNA damage-induced 'Lys-63'-linked polyubiquitination by PRPF19 mediates ATRIP recruitment to the RPA complex at sites of DNA damage and activation of ATR (PubMed:24332808). Ubiquitinated by RFWD3 at stalled replication forks in response to DNA damage: ubiquitination by RFWD3 does not lead to degradation by the proteasome and promotes removal of the RPA complex from stalled replication forks, promoting homologous recombination (PubMed:26474068).
Target Relevance information above includes information from UniProt accession: P15927
The UniProt Consortium

Data

benchmark-antibodies_anti-rpa32_antibody_6624_1.jpg
RPA32 antibody [12F3.3] detects RPA32 protein at DNA damage foci by immunofluorescent analysis.
Sample: 2mM hydroxyurea treated (right) or untreated (left) HeLa cells were fixed in ice-cold MeOH for 5 min.
Green: RPA32 protein stained by RPA32 antibody [12F3.3] (6624) diluted at 1:1000.
Blue: Hoechst 33342 staining.
Scale bar = 10 um.
benchmark-antibodies_anti-rpa32_antibody_6624_2.jpg
Detection of human RPA32 protein using RPA32 12F3.3 monoclonal antibody (6624) in HeLa (IR treated) whole cell extract. No radiation, 1hr, 2hr, and 4hr exposure corresponding to lanes 1 through 4 respectively.
benchmark-antibodies_anti-rpa32_antibody_6624_3.jpg
RPA32 antibody detects RPA32 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and blotted with RPA32 antibody (6624) diluted by 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.




pmidtitleauthorscitation

Protocols

relevant to this product
Western blot
IHC
ICC

Documents

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