Anti-RPA32/RPA2 antibody [9H8]

• Flow Cyt

Unknown

Flow Cytometry - Anti-RPA32/RPA2 antibody [9H8] (AB2175)

Overlay histogram showing HeLa cells stained with ab2175 (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 (ab2175, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [9H8] (AB2175)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining RPA32/RPA2 with ab2175. Staining was detected using DAB.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RPA32/RPA2 antibody [9H8] (AB2175)

Formalin-fixed, paraffin-embeded human tonsil tissue stained for RPA32/RPA2 using ab2175 at 1/50 dilution in immunohistochemical analysis. Antigen retrieval with citrate buffer pH 6.0

• Flow Cyt

PubMed

Flow Cytometry - Anti-RPA32/RPA2 antibody [9H8] (AB2175)

Anti-RPA/32/RPA2 antibody [9H8] (ab2175) is used to measure DNA Double-Stranded Breaks resection in U2OS cells +/- Camptotechin (CPT).

ab2175 is used at 1/200 dilution in 1xPBS + 0.2% Triton X-100 for 1 hour at 25°C.

Secondary antibody : anti-mouse Molecular Probes Alexa Fluor^® 488 Conjugated at 1/200 dilution.

This image is courtesy of an anonymous Abreview

• WB

CiteAb

Western blot - Anti-RPA32/RPA2 antibody [9H8] (AB2175)

Western Blotting using Anti-RPA32/RPA2 antibody [9H8], ab2175. Publication image from Gromak, N. et al., 2016, Nat Commun, 27725641. Legend direct from paper.

HRASV12 causes replication stress and genomic instability.(a) Top : DNA fibre labelling was performed in BJ-HRASV12 cells after RAS induction for 24, 48 and 72 h. Bottom : representative images of DNA fibres after 72 h RAS induction. (b) Distributions of replication fork speeds after RAS induction. N=3. (c) Median replication fork speeds after RAS induction. N=3. (d) Protein levels of phospho-S33 RPA32, RPA32 and GAPDH after RAS induction. (e) Protein levels of phospho-S345 CHK1, CHK1 and GAPDH after RAS induction. Hydroxyurea (HU, 2 mM for 24 h) was used as a positive control. (f) Representative images of γH2AX and 53BP1 foci after RAS induction for 96 h. (g) Percentages of cells containing more than 8 γH2AX or 53BP1 foci after RAS induction for 96 h. N=2. Asterisks compare with control. (h) Cell cycle distribution of 53BP1-positive cells after 96 h RAS induction as determined by co-staining with Cyclin A. Right panel : representative images; left panel : quantification. N=3. (i) Representative images of cells with micronuclei and percentages of cells containing micronuclei after RAS induction for 1–6 days. N=2. Asterisks compare with control. Means ±s.e.m. (bars) are shown. Student's t-test, *P<0.05 and **P<0.01. Scale bars, 10 µm.

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