Anti-PRPF8/Prp8 antibody [2834C1a]

• Flow Cyt

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Flow Cytometry - Anti-PRPF8/Prp8 antibody [2834C1a] (AB51366)

Overlay histogram showing HeLa cells stained with ab51366 (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 (ab51366, 2μg/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. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRPF8/Prp8 antibody [2834C1a] (AB51366)

IHC image of PRPF8/Prp8 staining in Human Normal Cervix formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab51366, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

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Western blot - Anti-PRPF8/Prp8 antibody [2834C1a] (AB51366)

All lanes:

Western blot - Anti-PRPF8/Prp8 antibody [2834C1a] (ab51366) at 1/100 dilution

All lanes:

HeLa whole cell lysate at 25 µg

Secondary

All lanes:

Anti mouse IgG antibody at 1/2500 dilution

Predicted band size: 274 kDa

Observed band size: 250 kDa

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• WB

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Western blot - Anti-PRPF8/Prp8 antibody [2834C1a] (AB51366)

All lanes:

Western blot - Anti-PRPF8/Prp8 antibody [2834C1a] (ab51366) at 1/100 dilution

All lanes:

NIH3T3 whole cell lysate at 25 µg

Secondary

All lanes:

Anti mouse IgG antibody at 1/2500 dilution

Predicted band size: 274 kDa

Observed band size: 250 kDa

false

• WB

CiteAb

Western blot - Anti-PRPF8/Prp8 antibody [2834C1a] (AB51366)

PRPF8/Prp8 western blot using anti-PRPF8/Prp8 antibody [2834C1a] ab51366. Publication image and figure legend from Pederiva, C., Bohm, S., et al., 2016, Cell Death Differ, PubMed 27315300.

ab51366 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 ab51366 please see the product overview.

When splicing-related factors are depleted, overexpression of RNF8 restores repair of DNA double-strand breaks. (a) Western blot of SF3B1, RBMX, PRPF8 and RNF8 in U2OS cells treated for 48 h with the siRNAs indicated. β-Actin was used as a loading control. (b) U2OS cells were transfected with the siRNAs indicated for 48 h, exposed to IR (6 Gy), and 1 h later, immunostained for γH2AX and conjugated ubiquitin (with the FK2 antibody). (c) U2OS cells were treated with the siRNAs indicated for 40 h, followed by transfection with either GFP-RNF8 or GFP-Empty plasmid for 7 h; exposure to IR (6 Gy) and fixation 1 h later and immunostaining for conjugated ubiquitin. The white numbers indicate the percentage of 100 transfected cells, i.e, green cells whose nuclei contained >10 IR-induced foci. Means±S.D. are shown, n=3. *P<0.05, as determined by a non-paired two-tailed Student's t-test. (d) Schematic model of how splicing regulates repair of DNA double-strand breaks

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