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AB325317

Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59]

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Rabbit Recombinant Monoclonal RPA32/RPA2 phospho S4 antibody. Suitable for Dot, ICC/IF, WB and reacts with Synthetic peptide - Human, Human, Mouse samples.
6 Images
Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) and 293T treated with camptothecin (1µM) for 3h labelling RPA32/RPA2 (phospho S4) with ab325317 at 1/500 (1.018 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing increased nuclear staining in 293T cell line treated with camptothecin (1µM) for 3 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab325317 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) and NIH/3T3 treated with camptothecin (1µM) for 3h labelling RPA32/RPA2 (phospho S4) with ab325317 at 1/500 (1.018 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing increased nuclear staining in NIH/3T3 cell line treated with camptothecin (1µM) for 3 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab325317 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)
  • WB

Supplier Data

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight of RPA32/RPA2 is upshifted in response to Etoposide treatment (PMID : 21731742).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-RPA32/RPA2 antibody (ab240637) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (ab325317) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) fresh whole cell lysate (untreated membrane) at 50 µg

Lane 2:

HeLa treated with 100µM Etoposide for 4h fresh whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Untreated HeLa fresh whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 4:

HeLa treated with 100µM Etoposide for 4h fresh whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 32 kDa,30 kDa,36 kDa

false

Exposure time: 26s

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)
  • WB

Supplier Data

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight of RPA32/RPA2 is upshifted in response to camptothecin treatment (PMID : 21731742).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-RPA32/RPA2 antibody (ab240637) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (ab325317) at 1/1000 dilution

Lane 1:

Untreated 293T (human embryonic kidney epithelial cell) fresh whole cell lysate (untreated membrane) at 50 µg

Lane 2:

293T treated with 1µM camptothecin for 3h fresh whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Untreated 293T (human embryonic kidney epithelial cell) fresh whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 4:

293T treated with 1µM camptothecin for 3h fresh whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 32 kDa,30 kDa,36 kDa

false

Exposure time: 15s

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)
  • WB

Supplier Data

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight of RPA32/RPA2 is upshifted in response to camptothecin treatment (PMID : 21731742).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-RPA32/RPA2 antibody (ab240637) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (ab325317) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast) fresh whole cell lysate (untreated membrane) at 50 µg

Lane 2:

NIH/3T3 treated with 1µM camptothecin for 3h fresh whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Untreated NIH/3T3 fresh whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 4:

NIH/3T3 treated with 1µM camptothecin for 3h fresh whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 32 kDa,30 kDa,36 kDa

false

Exposure time: 26s

Dot Blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)
  • Dot

Supplier Data

Dot Blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (AB325317)

Dot blot analysis of RPA32/RPA2 (phospho S4) using ab325317 at 1 : 1000 dilution (0.509 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human RPA32/RPA2 (phospho S4+S8) peptide a
Lane2 : Human RPA32/RPA2 (phospho S4) peptide b
Lane3 : Human RPA32/RPA2 (phospho S8) peptide c
Lane4 : Human RPA32/RPA2 non-phospho peptide d
Lane5 : Human RPA32/RPA2 (phospho S11+S12+S13) peptide e

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Dot Blot - Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] (ab325317) at 1/1000 dilution

Lane 1:

Human RPA32/RPA2 (phospho S4+S8) peptide a

Lane 2:

Human RPA32/RPA2 (phospho S4) peptide b

Lane 3:

Human RPA32/RPA2 (phospho S8) peptide c

Lane 4:

Human RPA32/RPA2 non-phospho peptide d

Lane 5:

Human RPA32/RPA2 (phospho S11+S12+S13) peptide e

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

  • Carrier free

    Anti-RPA32/RPA2 (phospho S4) antibody [EPR29611-59] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR29611-59

Isotype

IgG

Carrier free

No

Reacts with

Human, Mouse

Applications

WB, ICC/IF, Dot

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Synthetic peptide - Human": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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Product details

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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Shipped at conditions
Conditional Ambient
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
Avoid freeze / thaw cycle
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

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. Thereby, it plays an essential role both in DNA replication and the cellular response to DNA damage. 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. It is required for the recruitment of the DNA double-strand break repair factors RAD51 and RAD52 to chromatin in response to DNA damage. 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. Also plays a role in base excision repair (BER) probably through interaction with UNG. Also recruits SMARCAL1/HARP, which is involved in replication fork restart, to sites of DNA damage. May also play a role in telomere maintenance. RPA stimulates 5'-3' helicase activity of BRIP1/FANCJ (PubMed : 17596542).
See full target information RPA2 phospho S4

Alternative Names

REPA2, RPA32, RPA34, Replication protein A 32 kDa subunit, RP-A p32, Replication factor A protein 2, Replication protein A 34 kDa subunit, RF-A protein 2, RP-A p34, Rpa34, Replication protein A 32 kDa subunit, RP-A p32, Replication factor A protein 2, Replication protein A 34 kDa subunit, RF-A protein 2, RP-A p34
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com