Product: Phospho-RPA32/RPA2 (Thr21) Antibody
Catalog: AF3183
Description: Rabbit polyclonal antibody to Phospho-RPA32/RPA2 (Thr21)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse
Prediction: Pig, Zebrafish, Bovine, Sheep, Dog, Xenopus
Mol.Wt.: 35kDa; 29kD(Calculated).
Uniprot: P15927
RRID: AB_2834615

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse
Prediction:
Pig(100%), Zebrafish(88%), Bovine(100%), Sheep(100%), Dog(100%), Xenopus(83%)
Clonality:
Polyclonal
Specificity:
Phospho-RPA32/RPA2 (Thr21) Antibody detects endogenous levels of RPA32/RPA2 only when phosphorylated at Threonine 21.
RRID:
AB_2834615
Cite Format: Affinity Biosciences Cat# AF3183, RRID:AB_2834615.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

60S acidic ribosomal protein P1; AA409079; AI325195; AU020965; ik:tdsubc_2g1; M(2)21C; MGC137236; OTTHUMP00000004008; p32; p34; RCJMB04_6d17 replication protein A2, 32kDa; REPA2; Replication factor A protein 2; Replication protein A 32 kDa subunit; Replication protein A 32kDa subunit; Replication protein A 34 kDa subunit; Replication protein A; Replication Protein A2 (32kDa); Replication protein A2; Replication protein A2, 32kDa; RF-A protein 2; Rf-A2; RFA; RFA2_HUMAN; RP-A p32; RP-A p34; RP21C; RPA 2; RPA 32; RPA; Rpa2; RPA32; RPA34; RpLP1; RpP2; xx:tdsubc_2g1; zgc:109822;

Immunogens

Immunogen:

A synthesized peptide derived from human RPA32/RPA2 around the phosphorylation site of Thr21.

Uniprot:
Gene(ID):
Description:
RFA2 is absolutely required for simian virus 40 DNA replication in vitro. It participates in a very early step in initiation. A single-stranded DNA-binding protein.
Sequence:
MWNSGFESYGSSSYGGAGGYTQSPGGFGSPAPSQAEKKSRARAQHIVPCTISQLLSATLVDEVFRIGNVEISQVTIVGIIRHAEKAPTNIVYKIDDMTAAPMDVRQWVDTDDTSSENTVVPPETYVKVAGHLRSFQNKKSLVAFKIMPLEDMNEFTTHILEVINAHMVLSKANSQPSAGRAPISNPGMSEAGNFGGNSFMPANGLTVAQNQVLNLIKACPRPEGLNFQDLKNQLKHMSVSSIKQAVDFLSNEGHIYSTVDDDHFKSTDAE

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Zebrafish
88
Xenopus
83
Horse
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

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. Plays also 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.

PTMs:

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. 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.

Subcellular Location:

Nucleus. Nucleus>PML body.
Note: Redistributes to discrete nuclear foci upon DNA damage in an ATR-dependent manner.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Family&Domains:

Belongs to the replication factor A protein 2 family.

Research Fields

· Genetic Information Processing > Replication and repair > DNA replication.

· Genetic Information Processing > Replication and repair > Nucleotide excision repair.

· Genetic Information Processing > Replication and repair > Mismatch repair.

· Genetic Information Processing > Replication and repair > Homologous recombination.

· Genetic Information Processing > Replication and repair > Fanconi anemia pathway.

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