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AB325757

Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free

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Rabbit Recombinant Monoclonal RPA32/RPA2 phospho S33 antibody. Carrier free. Suitable for Dot, ICC/IF, WB and reacts with Synthetic peptide - Human, Human, Mouse samples.

View Alternative Names

REPA2, RPA32, RPA34, RPA2, 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

6 Images
Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)

This data was developed using ab325750, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell), HeLa treated with Etoposide(100uM) for 16hours and HeLa treated with Etoposide(100uM) for 16hours then Alkaline Phosphatase treatment 37℃ for 1 hour cells labelling RPA32/RPA2 (phospho S33) with ab325750 at 1/250 (2.06 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear staining in HeLa cells treated with Etoposide(100uM) for 16hours and weak staining in HeLa cells and HeLa cells treated with Etoposide(100uM) for 16hours then Alkaline Phosphatase treatment 37℃ for 1 hour(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).

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)

This data was developed using ab325750, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast), NIH/3T3 treated with Camptothecin/CPT(1uM) for 3hours and NIH/3T3 treated with Camptothecin/CPT(1uM) for 3hours then Alkaline Phosphatase treatment 37℃ for 1 hour cells labelling RPA32/RPA2 (phospho S33) with ab325750 at 1/250 (2.06 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear staining in NIH/3T3 cells treated with Camptothecin/CPT(1uM) for 3hours and weak staining in NIH/3T3 cells and NIH/3T3 cells treated with Camptothecin/CPT(1uM) for 3hours then Alkaline Phosphatase treatment 37℃ for 1 hour(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).

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)
  • WB

Lab

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)

This data was developed using ab325750, the same antibody clone in a different buffer formulation.

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

The molecular weight of RPA32/RPA2 is upshifted in response to Etoposide treatment (PMID : 17928296).

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) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] (<a href='/en-us/products/primary-antibodies/rpa32-rpa2-phospho-s33-antibody-epr29612-551-ab325750'>ab325750</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa treated with 100 μM Etoposide for 16 hours fresh whole cell lysate 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,36 kDa,30 kDa

false

Exposure time: 180s

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)
  • WB

Lab

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)

This data was developed using ab325750, the same antibody clone in a different buffer formulation.

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

The molecular weight of RPA32/RPA2 is upshifted in response to camptothecin treatment (PMID : 17928296).

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) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] (<a href='/en-us/products/primary-antibodies/rpa32-rpa2-phospho-s33-antibody-epr29612-551-ab325750'>ab325750</a>) 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 1 μM camptothecin for 3 h 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 1 μM camptothecin for 3 h 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,36 kDa,30 kDa

false

Exposure time: 180s

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)
  • WB

Lab

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)

This data was developed using ab325750, the same antibody clone in a different buffer formulation.

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

The molecular weight of RPA32/RPA2 is upshifted in response to camptothecin treatment (PMID : 17928296).

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.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] (<a href='/en-us/products/primary-antibodies/rpa32-rpa2-phospho-s33-antibody-epr29612-551-ab325750'>ab325750</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NIH/3T3 treated with 1 μM camptothecin for 3 h fresh whole cell lysate 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,36 kDa,30 kDa

true

Exposure time: 70s

Dot Blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)
  • Dot

Lab

Dot Blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] - BSA and Azide free (AB325757)

This data was developed using ab325750, the same antibody clone in a different buffer formulation.

Dot blot analysis of RPA32/RPA2 (phospho S33) using ab325750 at 1 : 1000 (0.515 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : Human RPA32/RPA2 (phospho S33) peptide a

Lane 2 : Human RPA32/RPA2 (phospho S33) peptide b

Lane 3 : Human RPA32/RPA2 (non-phospho) peptide

Lane 4 : Human RPA32/RPA2 (phospho S39) peptide

Exposure time : 180 seconds.

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

All lanes:

Dot Blot - Anti-RPA32/RPA2 (phospho S33) antibody [EPR29612-551] (<a href='/en-us/products/primary-antibodies/rpa32-rpa2-phospho-s33-antibody-epr29612-551-ab325750'>ab325750</a>) at 1/1000 dilution

Lane 1:

Human RPA32/RPA2 (phospho S33) peptide a

Lane 2:

Human RPA32/RPA2 (phospho S33) peptide b

Lane 3:

Human RPA32/RPA2 (non-phospho) peptide

Lane 4:

Human RPA32/RPA2 (phospho S39) peptide

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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR29612-551

Isotype

IgG

Carrier free

Yes

Reacts with

Human, Mouse

Applications

Dot, ICC/IF, WB

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"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "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": "", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-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": "", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-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": "", "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": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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Product details

ab325757 is the carrier-free version of ab325750

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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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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 S33
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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