Name: Anti-RPA70 antibody [EPR3472]
SKU: ab79398
Rating: 5 (6 reviews)

Anti-RPA70 antibody [EPR3472] (ab79398) is a rabbit monoclonal antibody detecting RPA70 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human.

- Biophysical QC for unrivalled batch-batch consistency

- Over 70 publications

- Trusted since 2009

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Images

Western blot - Anti-RPA70 antibody [EPR3472] (AB79398), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes See IHC antigen retrieval protocols. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/20	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes For unpurified use at 1/2000 - 1/5000.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/80	Notes For unpurified use at 1/50.Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

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Target data

See full target information RPA1

Function

As part of the heterotrimeric replication protein A complex (RPA/RP-A), binds and stabilizes single-stranded DNA intermediates that form during DNA replication or upon DNA stress. It prevents their reannealing and in parallel, recruits and activates different proteins and complexes involved in DNA metabolism (PubMed:17596542, PubMed:27723717, PubMed:27723720). Thereby, it plays an essential role both in DNA replication and the cellular response to DNA damage (PubMed:9430682). In the cellular response to DNA damage, the RPA complex controls DNA repair and DNA damage checkpoint activation. Through recruitment of ATRIP activates the ATR kinase a master regulator of the DNA damage response (PubMed:24332808). It is required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin in response to DNA damage (PubMed:17765923). Also recruits to sites of DNA damage proteins like XPA and XPG that are involved in nucleotide excision repair and is required for this mechanism of DNA repair (PubMed:7697716). Also plays a role in base excision repair (BER) probably through interaction with UNG (PubMed:9765279). Also recruits SMARCAL1/HARP, which is involved in replication fork restart, to sites of DNA damage. Plays a role in telomere maintenance (PubMed:17959650, PubMed:34767620). As part of the alternative replication protein A complex, aRPA, binds single-stranded DNA and probably plays a role in DNA repair. Compared to the RPA2-containing, canonical RPA complex, may not support chromosomal DNA replication and cell cycle progression through S-phase. The aRPA may not promote efficient priming by DNA polymerase alpha but could support DNA synthesis by polymerase delta in presence of PCNA and replication factor C (RFC), the dual incision/excision reaction of nucleotide excision repair and RAD51-dependent strand exchange (PubMed:19996105). RPA stimulates 5'-3' helicase activity of the BRIP1/FANCJ (PubMed:17596542).

Alternative names

REPA1, RPA70, RPA1, Replication protein A 70 kDa DNA-binding subunit, RP-A p70, Replication factor A protein 1, Single-stranded DNA-binding protein, RF-A protein 1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR3472
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

What is this antibody validated in?

Anti-RPA70 antibody [EPR3472] (ab79398) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human samples.

What is the molecular weight of RPA70?

Anti-RPA70 [EPR3472] (ab79398) specifically detects a band for RPA70 (UniProt: P27694) at a molecular weight of 70kDa.

Trusted by the scientific community

Anti-RPA70 [EPR3472] (ab79398) was first used in a scientific publication in 2009 and has been cited over 70 times in peer-reviewed journals.

Reviewed by scientists

Anti-RPA70 [EPR3472] (ab79398) has over 5 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR3472] also available for your convenience:
ab79398, Alexa Fluor® 488 - ab199097, Alexa Fluor® 647 - ab199240, Carrier free - ab239890, PE - ab303077, APC - ab303078, HRP - ab303079, Alkaline Phosphatase - ab308863, Alexa Fluor® 594 - ab310589, Alexa Fluor® 555 - ab312119, Alexa Fluor® 568 - ab312603, Alexa Fluor® 750 - ab321735

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-RPA70 antibody [EPR3472] (ab79398) at 1/4000 dilution
Lane 1: HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg
Lane 2: HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 68 kDa
Observed band size: 70 kDa
Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)
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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/200) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).
Immunoprecipitation - Anti-RPA70 antibody [EPR3472] (ab79398)
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.
All lanes: Immunoprecipitation - Anti-RPA70 antibody [EPR3472] (ab79398)
Predicted band size: 68 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 cervix carcinoma tissue labeling RPA70 with purified ab79398 at 1/100.
Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) 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.
Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)
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.
This image is courtesy of a customer review submitted by Remi Buisson.
Immunocytochemistry/ Immunofluorescence - Anti-RPA70 antibody [EPR3472] (ab79398)
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.
Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-RPA70 antibody [EPR3472] (ab79398) at 1/1000 dilution
All lanes: A549 (Human lung carcinoma cell line) cell lysate at 20 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 68 kDa
Observed band size: 70 kDa
Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
All lanes: Western blot - Anti-RPA70 antibody [EPR3472] (ab79398) at 1/5000 dilution
Lane 1: A549 (Human lung carcinoma cell line) cell lysate at 10 µg
Lane 2: HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg
Secondary
All lanes: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 68 kDa
Observed band size: 70 kDa
Flow Cytometry (Intracellular) - Anti-RPA70 antibody [EPR3472] (ab79398)
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) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.
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.
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-RPA70 antibody [EPR3472] (ab79398)
RPA70 western blot using anti-RPA70 antibody [EPR3472] ab79398. Publication image and figure legend from Pedersen, H., Anne Adanma Obara, E., et al., 2020, Int J Mol Sci, PubMed 32111042.

ab79398 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab79398 please see the product overview.
RPA expression is crucial for the maintenance of glioblastoma cancer stem-like cells. (A) Immunoblot analysis of RPA70, RPA32 and RPA14 in a panel of GBM primary cell lines (4121, G01, G06, G40, G07, G16 and G20) and two normal human astrocyte cell lines (NHA33, NHA26). β-Actin was used as a loading control. (B) Immunoblot analysis of RPA70, RPA32 and RPA14 expression in matched pairs of glioblastoma cancer stem-like cells (GSCs) and differentiated GBM cells (DGCs) isolated from 4121, G01, G06 and G40 primary GBM cell lines. α-Tubulin or Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (C) Immunoblot analysis validating knockdown efficiencies of lentiviral shRNAs targeting RPA70, RPA32 and RPA14 in G01-GSCs. α-Tubulin was used as a loading control. (D) CellTiter-Glo luminescent viability assay (Promega) of G01-GSCs transduced with lentiviral shRNA targeting RPA70, RPA32 or RPA14 assessed 72 h after virus wash. Data are presented as mean ± SD. Statistical significance was tested using a two-way ANOVA with post-hoc Sidak’s multiple comparisons test, where *** p < 0.001. N = 3. (E) CellTiter-Glo luminescent viability assay (Promega) of G01-GSCs transduced with lentiviral shRNA targeting RPA70, RPA32 or RPA14 alone or 24 h after irradiation (3 Gy; COMBO) or sham-irradiated and assessed for viability 72 h later. (E) Extreme limiting dilution assay (ELDA) of G01-GSCs transduced with lentiviral shRNA targeting of RPA70 (2 independent shRNAs) shows significantly impaired self-renewal of G01-GSCs after 10 days. Data are presented as mean ± SD. Statistical significance was tested using a two-way ANOVA with post-hoc Sidak’s multiple comparisons test, where * p < 0.05, ** p < 0.01, *** p < 0.001. N = 3.