Recombinant Anti-RPA70 antibody [EPR3472] (ab79398)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3472] to RPA70
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-RPA70 antibody [EPR3472]
See all RPA70 primary antibodies -
Description
Rabbit monoclonal [EPR3472] to RPA70 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3472 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab79398 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) |
1/80.
For unpurified use at 1/50.ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (2) |
1/100 - 1/250.
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WB | (1) |
1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
For unpurified use at 1/2000 - 1/5000. |
IP | (1) |
1/10 - 1/20.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/80. For unpurified use at 1/50.ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa). For unpurified use at 1/2000 - 1/5000. |
IP
1/10 - 1/20. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
Plays an essential role in several cellular processes in DNA metabolism including replication, recombination and DNA repair. Binds and subsequently stabilizes single-stranded DNA intermediates and thus prevents complementary DNA from reannealing.
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. -
Sequence similarities
Belongs to the replication factor A protein 1 family. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Sumoylated on lysine residues Lys-449 and Lys-577, with Lys-449 being the major site. Sumoylation promotes recruitment of RAD51 to the DNA damage foci to initiate DNA repair through homologous recombinaison. Desumoylated by SENP6. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 6117 Human
- Omim: 179835 Human
- SwissProt: P27694 Human
- Unigene: 461925 Human
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Alternative names
- Dmrpa1 antibody
- Drosophila Replication Protein A antibody
- DRPA antibody
see all
Images
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All lanes : Anti-RPA70 antibody [EPR3472] (ab79398) at 1/4000 dilution (purified)
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Unpurified ab79398 staining RPA70 in U-2 OS (Human bone osteosarcoma epithelial cell line) cells by ICC/IF (Immunocytochemistry/immunofluorescence).
Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 in PBS and blocked with 2% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/500 in PBS + 0.5% Tween-20) for 2 hours at 25°C. A Cy3®-conjugated goat anti-rabbit IgG monoclonal (1/250) was used as the secondary antibody.
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Immunocytochemistry/Immunofluorescence analysis of A549 (Human lung carcinoma cell line) cells labeling RPA70 with purified ab79398 at 1/200.
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/200) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling RPA70 with purified ab79398 at 1/100.
Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).
Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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ab79398 (purified) at 1/20 immunoprecipitating RPA70 in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10 µg)
Lane 2 (+): ab79398 + HeLa whole cell lysate (10 µg).
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab79398 in HeLa whole cell lysate.
For western blotting, an HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking/Dilution buffer: 5% NFDM/TBST.
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Intracellular Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling RPA70 with purified ab79398 at 1/80 (red).
Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.
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Anti-RPA70 antibody [EPR3472] (ab79398) at 1/1000 dilution (purified) + A549 (Human lung carcinoma cell line) cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-RPA70 antibody [EPR3472] (ab79398) at 1/5000 dilution (unpurified)
Lane 1 : A549 (Human lung carcinoma cell line) cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical squamous cell carcinoma labelling RPA70 with unpurified ab79398 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with unpurifiedab79398 (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 (unpurified ab79398, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C.
Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/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 (65)
ab79398 has been referenced in 65 publications.
- Osborne HC et al. Sesquiterpene Lactones Potentiate Olaparib-Induced DNA Damage in p53 Wildtype Cancer Cells. Int J Mol Sci 23:N/A (2022). PubMed: 35163037
- Luo H et al. HOTTIP-dependent R-loop formation regulates CTCF boundary activity and TAD integrity in leukemia. Mol Cell 82:833-851.e11 (2022). PubMed: 35180428
- McQuaid ME et al. Hypomorphic GINS3 variants alter DNA replication and cause Meier-Gorlin syndrome. JCI Insight 7:N/A (2022). PubMed: 35603789
- Mattoo AR & Jessup JM MCL-1 interacts with MOF and BID to regulate H4K16 acetylation and homologous recombination repair. Cell Biol Int 46:1196-1203 (2022). PubMed: 35661479
- Hussain M et al. A small-molecule Skp1 inhibitor elicits cell death by p53-dependent mechanism. iScience 25:104591 (2022). PubMed: 35789855