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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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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Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)This image is courtesy of an Abreview submitted by Remi Buisson
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) - Anti-RPA70 antibody [EPR3472] (ab79398)
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) - Anti-RPA70 antibody [EPR3472] (ab79398)
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