Anti-RPA32/RPA2 antibody [9H8] (ab2175)
- Product nameAnti-RPA32/RPA2 antibody [9H8]See all RPA32/RPA2 primary antibodies ...
- DescriptionMouse monoclonal [9H8] to RPA32/RPA2
- SpecificityThis antibody reacts with a 32-34 kDa protein known as Replication Protein A (RPA).
- Tested applicationsFlow Cyt, ChIP, WB, IP, IHC-P, IHC-Fr, ICC/IF more details
- Species reactivityReacts with: Human
Full length protein (Human).
- Positive control
- Storage instructionsStore at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
- Storage bufferPreservative: 0.05% Sodium Azide
Constituents: 1% BSA
- Concentration information loading...
- PurityIgG fraction
- Clonality Monoclonal
- Clone number9H8
- Light chain typekappa
- Research Areas
Our Abpromise guarantee covers the use of ab2175 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
|ChIP||ChIP: Use at an assay dependent concentration.|
|WB||WB: Use at an assay dependent dilution. Predicted molecular weight: 32 kDa.|
|IP||IP: Use at an assay dependent dilution.|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|IHC-Fr||IHC-Fr: Use at an assay dependent dilution. This antibody may be diluted to a titer of 1/50-1/100 in an ABC method. We suggest an incubation period of 30 minutes at room temperature.|
|ICC/IF||ICC/IF: Use at an assay dependent concentration. Used at a dilution of 1/300 for 2 hrs on mouse MEF cells (see Abreview for further details).|
- FunctionRequired for DNA recombination, repair and replication. The activity of RP-A is mediated by single-stranded DNA binding and protein interactions.
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.
modificationsPhosphorylated in a cell-cycle-dependent manner (from the S phase until mitosis). Phosphorylated by ATR upon DNA damage, which promotes its translocation to nuclear foci. Can be phosphorylated in vitro by PRKDC/DNA-PK in the presence of Ku and DNA, and by CDK1.
- Cellular localizationNucleus. Nucleus > PML body. Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage.
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Anti-RPA32/RPA2 antibody [9H8] images
ab2175 - immunohistochemistryFormalin fixed paraffin embedded human tonsil stained with RPA using ABC and AEC chromogen.
ab2175 at 1/200 staining human skin fibroblasts by ICC/IF. The cells were treated with 2mM hydroxyurea for 16 hours, formaldehyde fixed and permeabilized with 0.5% Triton X100. They were then incubated with the primary antibody for 1.5 hours at 37°C. A Cy3 ® conjugated sheep anti-mouse antibody was used as the secondary.
All lanes : Anti-RPA32/RPA2 antibody [9H8] (ab2175) at 5 µg/ml
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 : SK N BE (Human neuroblastoma) Whole Cell Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lysates/proteins at 10 µg per lane.
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size : 32 kDa
Observed band size : 32 kDa
Anti-RPA/32/RPA2 antibody [9H8] (ab2175) is used to measure DNA Double-Stranded Breaks resection in U2OS cells +/- Camptotechin (CPT).
ab2175 is used at 1/200 dilution in 1xPBS + 0.2% Triton X-100 for 1 hour at 25°C.
Secondary antibody: anti-mouse Molecular Probes Alexa Fluor® 488 Conjugated at 1/200 dilution.
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining RPA32/RPA2 with ab2175. Staining was detected using DAB.
Overlay histogram showing HeLa cells stained with ab2175 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2175, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-RPA32/RPA2 antibody [9H8] (ab2175)
This product has been referenced in:
- Bétous R et al. Identification and characterization of SMARCAL1 protein complexes. PLoS One 8:e63149 (2013). WB, IP ; Human . Read more (PubMed: 23671665) »
- Qin Z et al. DNA-damage tolerance mediated by PCNA*Ub fusions in human cells is dependent on Rev1 but not Pol?. Nucleic Acids Res 41:7356-69 (2013). ICC/IF ; Human . Read more (PubMed: 23761444) »