Recombinant Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2877Y] to RPA32/RPA2
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RPA32/RPA2 antibody [EPR2877Y]
See all RPA32/RPA2 primary antibodies -
Description
Rabbit monoclonal [EPR2877Y] to RPA32/RPA2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human RPA32/RPA2 (C terminal). The exact sequence is proprietary.
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Positive control
- IHC-P: Human breast carcinoma tissue, mouse and rat liver Tissue WB: HeLa whole cell lysate (ab150035), HUVEC lysate, NIH3T3 cell lysate, C6 cell lysate ICC/IF: C6, HeLa, NIH/3T3 cells. Flow Cyt (intra): C6, HeLa, NIH/3T3 cells. IP: HeLa cell lysate
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2877Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76420 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 (Intra) |
Use at an assay dependent concentration.
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WB |
1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).
For unpurified format use at 1/5000 - 1/10000. |
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IP |
1/20.
For unpurified format use at 1/40. |
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/50.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa). For unpurified format use at 1/5000 - 1/10000. |
IP
1/20. For unpurified format use at 1/40. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/50. |
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
- Entrez Gene: 19891 Mouse
- Entrez Gene: 59102 Rat
- Omim: 179836 Human
- SwissProt: P15927 Human
- SwissProt: Q62193 Mouse
- SwissProt: Q63528 Rat
- Unigene: 703070 Human
see all -
Alternative names
- 60S acidic ribosomal protein P1 antibody
- AA409079 antibody
- AI325195 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 29 kDa
Observed band size: 29 kDa -
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/50 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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All lanes : Anti-RPA32/RPA2 antibody [EPR2877Y] (ab76420) at 1/10000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : HUVEC lysate
Lane 3 : NIH 3T3 cell lysate
Lane 4 : C6 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/1000 dilution
Predicted band size: 29 kDa
Observed band size: 29 kDa -
Intracellular Flow Cytometry analysis of C6 (rat glial tumor glial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/200 dilution (0.58 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
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ab76420 (purified) at 1/20 dilution (0.5ug) immunoprecipitating RPA32/RPA2 in HeLa whole cell lysate. HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab76420 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76420 in HeLa whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 µg/ml). Heat mediated antigen retrieval was performed ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling RPA32/RPA2 with purified ab76420 at 1/100 dilution (1 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1:50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling RPA32/RPA2 with purified ab76420 at 1:50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1:50 (2.28 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling RPA32/RPA2 with purified ab76420 at 1/200 dilution (0.58 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling RPA32/RPA2 with purified ab76420 at 1/200 dilution (0.58 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab76420 (unpurified) at 1/250 dilution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (24)
ab76420 has been referenced in 24 publications.
- Abate NG & Hendzel MJ Heterogeneity of Organization of Subcompartments in DSB Repair Foci. Front Genet 13:887088 (2022). PubMed: 35923694
- Nore A et al. TOPOVIBL-REC114 interaction regulates meiotic DNA double-strand breaks. Nat Commun 13:7048 (2022). PubMed: 36396648
- Singh AK et al. Local DNA synthesis is critical for DNA repair during oocyte maturation. J Cell Sci 134:N/A (2021). PubMed: 34415018
- Yan S et al. ZGRF1 promotes end resection of DNA homologous recombination via forming complex with BRCA1/EXO1. Cell Death Discov 7:260 (2021). PubMed: 34552057
- Song X et al. Synergistic targeting of CHK1 and mTOR in MYC-driven tumors. Carcinogenesis 42:448-460 (2021). PubMed: 33206174