Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Orangutan (100%). Backed by our 100% Guarantee.

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all RPA2 Primary Antibodies »

Product Specifications

Immunogen

Immunogen was a phosphorylated synthetic peptide, which represented a portion of human replication protein A2, 32 kDa surrounding phosphorylated serine that corresponded to position 33 using the numbering given in entry NP_002937.1 (GeneID 6118).

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:33087072).

Modification

p Ser33

Specificity

NB100-544 is specific for human RPA-32 pSer33 protein.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

32 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for RPA2 [p Ser33] Antibody - BSA Free

Western Blot: RPA2 [p Ser33] Antibody [NB100-544]

Western Blot: RPA2 [p Ser33] Antibody [NB100-544]

Western Blot: RPA2 [p Ser33] Antibody [NB100-544] - Whole cell lysate (50 ug) from HeLa cells treated with 100 uM etoposide for 16 hours (+) or mock treated (-) cells. Antibody: Affinity purified rabbit anti-Phospho RPA32 (S33) antibody used for WB at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes. Lower panel: Rabbit anti-RPS3 antibody.
Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544]

Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544]

RPA2-[p-Ser33]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-544-img0004.jpg
Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544]

Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544]

Immunocytochemistry/Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544] - L.36 PDAC cells were treated two hours using 5 mM hydroxyurea to induce RPA32 Ser33 phosphorylation. The antibody produces clear nuclear staining of treated cells. ICC/IF image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544]

Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody [NB100-544]

RPA2-[p-Ser33]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-544-img0003.jpg
RPA2 [p Ser33] Antibody - BSA Free

Western Blot: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

The effects of ARID1A on the response to replication stress and DSB induced by HU. (A and B) Western blot analysis of DDR-related genes following the exposure to 2 mM HU (A) and 4 mM HU (B) for the indicated times in ARID1A KD or OE DMS273 cells. (C to H) Cell viability (C and F) and clonal formation (D, E, G, and H) assays following 2 mM HU (C to E) and 4 mM HU (F to H) in ARID1A KD or OE DMS273 cells. n ≥ 3 independent experiments. Data are shown as the mean +/- SEM; n ≥ 3 independent experiments. Statistical analysis was performed using a one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41049615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RPA2 [p Ser33] Antibody - BSA Free

Western Blot: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

ARID1A deficiency enhances JQ1 sensitivity and identifies BRD-K98645985 as a novel therapeutic candidate in SCLC. (A and B) Pearson correlation analysis of IC50 values of JQ1 from the GDSC1 dataset and the mRNA level of ARID1A (A) and MYC family gene (B). (C) Dose–response analysis of JQ1 treatment on cell viability upon ARID1A KD DMS273 and DMS53 cells. (D) Western blot analysis of DDR-related genes following the exposure to JQ1 at indicated concentrations in ARID1A KD DMS273 and DMS53 cells. (E) Tumor volume curves of DMS273 control cells (SCR) and ARID1A KD cells upon exposure to JQ1 in vivo. KD denotes shARID1A-2#. (F) Scatterplot representing tumor weights of DMS273 control cells (SCR) and ARID1A KD cells upon exposure to JQ1 at the endpoint of experiments in vivo. KD refers to shARID1A-2#. (G) Colony formation assays demonstrating the effect of BRD-K98645985 on the cytotoxicity of JQ1 in DMS273 cells. (H) Drug–response curves of BRD-K98645985 and JQ1 combination in DMS273 cells. Cells were treated with escalating doses for 72 h, with viability measured by CellTiter-Glo. (I) Bliss analysis (Combenefit software) showing synergistic effects. (J) Tumor growth curves in xenograft mice treated with JQ1, BRD-K98645985 (BRD), or their combination (JQ1 + BRD). (K) Scatterplot depicting tumor weights from xenograft mice at the endpoints of the experiments. (L) Representative images of xenograft tumors. Data are mean +/- SEM, and statistical analysis was performed using a one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (M) Proposed model depicting the function of ARID1A to modulate cell proliferation and genome stability through c-MYC and PARP1. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41049615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RPA2 [p Ser33] Antibody - BSA Free

Western Blot: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

ARID1A participates in DDR in SCLC through the c-MYC/PARP axis. (A and B) Western blot analysis of DDR-related gene in DMS273 and DMS53 cells following ARID1A KD (A) or OE (B). (C and D) Representative images of immunofluorescence staining for gamma -H2AX (C) and RAD51 (D) in DMS273 and DMS53 cells following ARID1A KD. Cells with more than 5 foci were considered positive. Quantification of gamma -H2AX /RAD51 fluorescence intensities from 3 independent experiments was shown as mean +/- SD. Statistical analysis was performed using 2-tailed unpaired Student’s t tests. ****P < 0.0001. (E to G) Western blot analysis of the indicated proteins following ARID1A (E), c-MYC (F), or PARP1 (G) overexpression in ARID1A KD DMS273 and DMS53 cells. (H and I) Western blot analysis of PI3K downstream phospho-proteins in DMS273 and DMS53 cells following ARID1A KD (H) or OE (I). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41049615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RPA2 [p Ser33] Antibody - BSA Free

Western Blot: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

The effects of ARID1A on the response to replication stress and DSB induced by HU. (A and B) Western blot analysis of DDR-related genes following the exposure to 2 mM HU (A) and 4 mM HU (B) for the indicated times in ARID1A KD or OE DMS273 cells. (C to H) Cell viability (C and F) and clonal formation (D, E, G, and H) assays following 2 mM HU (C to E) and 4 mM HU (F to H) in ARID1A KD or OE DMS273 cells. n ≥ 3 independent experiments. Data are shown as the mean +/- SEM; n ≥ 3 independent experiments. Statistical analysis was performed using a one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41049615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RPA2 [p Ser33] Antibody - BSA Free

Western Blot: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

ARID1A participates in DDR in SCLC through the c-MYC/PARP axis. (A and B) Western blot analysis of DDR-related gene in DMS273 and DMS53 cells following ARID1A KD (A) or OE (B). (C and D) Representative images of immunofluorescence staining for gamma -H2AX (C) and RAD51 (D) in DMS273 and DMS53 cells following ARID1A KD. Cells with more than 5 foci were considered positive. Quantification of gamma -H2AX /RAD51 fluorescence intensities from 3 independent experiments was shown as mean +/- SD. Statistical analysis was performed using 2-tailed unpaired Student’s t tests. ****P < 0.0001. (E to G) Western blot analysis of the indicated proteins following ARID1A (E), c-MYC (F), or PARP1 (G) overexpression in ARID1A KD DMS273 and DMS53 cells. (H and I) Western blot analysis of PI3K downstream phospho-proteins in DMS273 and DMS53 cells following ARID1A KD (H) or OE (I). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41049615), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RPA2 [p Ser33] Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

Active DNA end processing in late micronuclei compartments. a: Detection and quantification of RPA32 phosphorylation (pSerine 33) and ssDNA by specific antibodies in MN compartments of OVCAR-8 cells 36 h after 2-Gy irradiation. Note the nuclear signals have reduced while those in MN were intense. n = 3 biological repeats. Error bars = s.d. **P < 0.01; ***P < 0.005 (t-test). b: Immunostaining and enumeration of phosphorylated NBS1-pSerine 343, BRCA1-pSerine 1524 and TP53-pSerine 15 in MN compartments. **P < 0.01; ***P < 0.005 (t-test). c: Representative images (Upper panel) and quantification of RPA signal in MN compartments (Lower left) or RPA foci in nuclei (lower right) of post-IR OVCAR-8 cells after indicated treatments. Pro-resection genes (MRE11, BRCA1 and WDR70) were knockdown by specific siRNA. Percentage of positive RPA32-pS33 MN population was calculated after 36 h of IR treatment Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29661159), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
RPA2 [p Ser33] Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: RPA2 [p Ser33] Antibody - BSA Free [NB100-544] -

Active DNA end processing in late micronuclei compartments. a: Detection and quantification of RPA32 phosphorylation (pSerine 33) and ssDNA by specific antibodies in MN compartments of OVCAR-8 cells 36 h after 2-Gy irradiation. Note the nuclear signals have reduced while those in MN were intense. n = 3 biological repeats. Error bars = s.d. **P < 0.01; ***P < 0.005 (t-test). b: Immunostaining and enumeration of phosphorylated NBS1-pSerine 343, BRCA1-pSerine 1524 and TP53-pSerine 15 in MN compartments. **P < 0.01; ***P < 0.005 (t-test). c: Representative images (Upper panel) and quantification of RPA signal in MN compartments (Lower left) or RPA foci in nuclei (lower right) of post-IR OVCAR-8 cells after indicated treatments. Pro-resection genes (MRE11, BRCA1 and WDR70) were knockdown by specific siRNA. Percentage of positive RPA32-pS33 MN population was calculated after 36 h of IR treatment Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29661159), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for RPA2 [p Ser33] Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500 - 1:2500

Simple Western

1:25

Western Blot

1:1000 - 1:5000
Application Notes
Numerous bands representing proteins that migrate more slowly than 60 to 70 kDA have been observed during WB. Under some conditions, at least two of these may be more intense than the band that represents phospho RPA32-S33. ICC/IF reactivity reported in (PMID: 27723720).
See Simple Western Antibody Database for Simple Western validation: tested in Human squamous carcinoma cells; antibody dilution of 1:25; separated by charge

Reviewed Applications

Read 1 review rated 5 using NB100-544 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Citrate/Phosphate (pH 7.0 - 8.0)

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: RPA2

Replication Protein A (RPA) is a single stranded DNA binding protein. Human RPA is a heterotrimeric protein containing subunits of 14, 32 and 70kDa. This protein complex is highly conserved in eukaryotes and is essential in DNA replication, homologous recombination and nucleotide excision repair. RPA is a target for DNA dependent protein kinase (DNA-PK), Ataxia Telangiectasia Mutated gene (ATM) and a cyclin dependent protein kinase. The middle subunit, RPA 32, is phosphorylated during the S-phase of the cell cycle in response to DNA damage, and during apoptosis of Jurkat T-lymphocytes.

Long Name

Replication Protein A2, 32kDa

Alternate Names

REPA2, RPA32

Entrez Gene IDs

6118 (Human)

Gene Symbol

RPA2

UniProt

P15927 (Human)

Additional RPA2 Products

Product Documents for RPA2 [p Ser33] Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for RPA2 [p Ser33] Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for RPA2 [p Ser33] Antibody - BSA Free

Customer Reviews for RPA2 [p Ser33] Antibody - BSA Free (1)

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  • RPA2 [p Ser33] Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: L3.6 Pancreatic cancer cell line
    Species: Human
    Verified Customer | Posted 08/04/2020
    L.36 PDAC cells were treated during two hours using 5mM hydroxyurea to induce RPA32 Ser33 phosphorylation. The antibody produces clear nucear staining of treated cells.
    Cells were treated with Hydroxyurea 5mM during 2 hours to induce replication stress and RPA32 phosphorylation. RPA32 P-Ser33 primary antibody dilution 1:2500 Secondary antibody: Rabbit Alexa 555 conjugated 1:500 DAPI counterstaining on cell nuclei
    RPA2 [p Ser33] Antibody - BSA Free NB100-544

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