Anti-RPA32/RPA2 antibody [9H8] (ab2175)
Key features and details
- Mouse monoclonal [9H8] to RPA32/RPA2
- Suitable for: Flow Cyt, WB, IHC-P
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-RPA32/RPA2 antibody [9H8]
See all RPA32/RPA2 primary antibodies -
Description
Mouse monoclonal [9H8] to RPA32/RPA2 -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Full length native protein (purified) corresponding to Human RPA32/RPA2. BALB/C mice were injected with purified human RPA protein
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Positive control
- WB: TE 671, SK N BE and HeLa whole cell lysates. IHC-P: Human tonsil tissue. Flow Cyt: HeLa cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.3
Preservative: 0.05% Sodium azide
Constituent: 1% BSA -
Concentration information loading...
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Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
9H8 -
Myeloma
unknown -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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Flow Cyt | (1) |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB | (2) |
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Required 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. -
Post-translational
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 localization
Nucleus. Nucleus > PML body. Also present in PML nuclear bodies. Redistributes to discrete nuclear foci upon DNA damage. - Information by UniProt
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Database links
- Entrez Gene: 6118 Human
- Omim: 179836 Human
- SwissProt: P15927 Human
- Unigene: 703070 Human
- Unigene: 79411 Human
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Alternative names
- 60S acidic ribosomal protein P1 antibody
- AA409079 antibody
- AI325195 antibody
see all
Images
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Formalin-fixed, paraffin-embeded human tonsil tissue stained for RPA32/RPA2 using ab2175 at 1/50 dilution in immunohistochemical analysis. Antigen retrieval with citrate buffer pH 6.0
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining RPA32/RPA2 with ab2175. Staining was detected using DAB.
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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.
Secondary
All lanes : 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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (223)
ab2175 has been referenced in 223 publications.
- Wang YC et al. Arginine shortage induces replication stress and confers genotoxic resistance by inhibiting histone H4 translation and promoting PCNA ubiquitination. Cell Rep 42:112296 (2023). PubMed: 36961817
- Guervilly JH et al. SLX4 dampens MutSα-dependent mismatch repair. Nucleic Acids Res 50:2667-2680 (2022). PubMed: 35166826
- Serafim RB et al. PIMREG expression level predicts glioblastoma patient survival and affects temozolomide resistance and DNA damage response. Biochim Biophys Acta Mol Basis Dis 1868:166382 (2022). PubMed: 35301087
- Mehta KPM et al. CHK1 phosphorylates PRIMPOL to promote replication stress tolerance. Sci Adv 8:eabm0314 (2022). PubMed: 35353580
- Egger T et al. A clinically relevant heterozygous ATR mutation sensitizes colorectal cancer cells to replication stress. Sci Rep 12:5422 (2022). PubMed: 35361811