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RPA32/RPA2 Antibody (YA679)

2 Cited Publications
Customer Review

Based on 2 publication(s) in Google Scholar

RPA32/RPA2 Antibody (YA679) is a Mouse-derived and non-conjugated IgG2b monoclonal antibody, targeting to RPA32/RPA2.

For research use only. We do not sell to patients.
  • Host:

    Mouse

  • Isotype:

    IgG2b

  • Application:

    WB, IHC-P, ICC/IF, IP

  • Reactivity :

    Human, Mouse, Rat

  • Formulation:

    Supplied in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide, pH 7.3.

  • Conjugation:
    Non-conjugated
ALL WB ICC
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • RPA32/RPA2 Antibody (YA679) Product Image
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Applications

Application
WB Info
WB: Western Blot
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
IP Info
IP: Immunoprecipitation
Dilution Ratio 1:500-1:1000 1:50-1:100 1:50-1:200 1:20

Product Details

Description

RPA32/RPA2 Antibody (YA679) is a Mouse-derived and non-conjugated IgG2b monoclonal antibody, targeting to RPA32/RPA2.

  • Host Mouse
  • Clonality Monoclonal
  • Species Reactivity
    Human, Mouse, Rat
  • Observed Molecular Weight
    Observed band size: 32 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 29 kDa
Species Reactivity Database
Immunogen

Purified recombinant human RPA32/RPA2 protein fragments expressed in E.coli.

Sensitivity

Endogenous

Purification

Protein A affinity purified.

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG2b

RRID

AB_3102780

Product Properties

  • Appearance

    Liquid

  • Formulation

    Supplied in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide, pH 7.3.

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
    Western blot analysis of extracts from Hela (lane 2(20μg), HUVEC (lane 3(20μg), MCF-7 (lane 4(20μg), using RPA32 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
    Immunocytochemistry analysis of Hela cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
    Immunocytochemistry analysis of NIH3T3 cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Background

  • Function

    RPA32/RPA2 is an 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. RPA stimulates 5'-3' helicase activity of BRIP1/FANCJ[1].

  • Subcellular Localization

    Nucleus; Nucleus, PML body

  • Expression


    Induction:Translationally up-regulated in response to DNA damage (at protein level)

  • Isoforms & Post-Translational Modification

    P15927 has 3 isomers: P15927-1: 29247 Da (predicted); P15927-2: 30156 Da (predicted); P15927-3: 38810 Da (predicted).
    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)

  • Subunit

    Component of the replication protein A complex (RPA/RP-A), a heterotrimeric complex composed of RPA1, RPA2 and RPA3 (PubMed:10449415, PubMed:19116208, PubMed:2406247). 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, PubMed:24910198). Interacts with RAD52 and XPA; those interactions are direct and associate RAD52 and XPA to the RPA complex (PubMed:11081631, PubMed:17765923, PubMed:7700386, PubMed:8702565). Interacts with FBH1 (PubMed:23319600). Interacts with ETAA1; the interaction is direct and promotes ETAA1 recruitment at stalled replication forks (PubMed:27601467, PubMed:27723717, PubMed:27723720). 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). Interacts with BRIP1/FANCJ via the RPA1 subunit; following DNA damage they colocalize in foci in the nucleus (PubMed:17596542). Interacts with nuclear UNG (isoform 2); this interaction mediates UNG recruitment to RPA-coated single-stranded DNA at stalled replication forks

  • SwissProt ID

    P15927

  • Gene ID
    6118 [NCBI]

RPA32/RPA2 Antibody (YA679) Related Classifications

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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedchemExpress Validation
     Western blot analysis of extracts from Hela (lane 2(20μg), HUVEC (lane 3(20μg), MCF-7 (lane 4(20μg), using RPA32 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

  • 1/16

    MedchemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • 1/16

    MedchemExpress Validation
     Immunocytochemistry analysis of NIH3T3 cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • RPA32/RPA2 Antibody (YA679) Product Image
    1/16
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • RPA32/RPA2 Antibody (YA679) Product Image
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
    1/1
    MedchemExpress Validation
     Western blot analysis of extracts from Hela (lane 2(20μg), HUVEC (lane 3(20μg), MCF-7 (lane 4(20μg), using RPA32 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
    1/1
    MedchemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
    1/1
    MedchemExpress Validation
     Immunocytochemistry analysis of NIH3T3 cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • Experimental Validation Results for RPA32/RPA2 Antibody (YA679)
  • RPA32/RPA2 Antibody (YA679) Product Image
    MedchemExpress Validation

    Western blot analysis of extracts from Hela (lane 2(20μg), HUVEC (lane 3(20μg), MCF-7 (lane 4(20μg), using RPA32 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

  • RPA32/RPA2 Antibody (YA679) Product Image
    MedchemExpress Validation

    Immunocytochemistry analysis of Hela cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • RPA32/RPA2 Antibody (YA679) Product Image
    MedchemExpress Validation

    Immunocytochemistry analysis of NIH3T3 cells labeling RPA32 with RPA32 Antibody (HY-P80975) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RPA32 Antibody (HY-P80975)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • RPA32/RPA2 Antibody (YA679) Product Image WB
  • RPA32/RPA2 Antibody (YA679) Product Image ICC
  • RPA32/RPA2 Antibody (YA679) Product Image ICC