Anti-Mre11 antibody [12D7] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

Immunocytochemical analysis of, 4% paraformaldehyde-fixed at RT for 15 min, HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Mre-11 (green) with ab214 at 1/200 dilution. Blue : Hoechst 33342 staining. Scale bar= 10 μm.

• Flow Cyt

Unknown

Flow Cytometry - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

Overlay histogram showing HeLa cells stained with ab214 (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 (ab214, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (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. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

This image was generated using the ascites version of the product.

• WB

Supplier Data

Western blot - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

Samples were separated by 7.5% SDS-PAGE.

All lanes:

Western blot - Anti-Mre11 antibody [12D7] - BSA and Azide free (ab214) at 1/1000 dilution

Lane 1:

293T whole cell extracts at 30 µg

Lane 2:

A431 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-mouse IgG antibody

false

• WB

Supplier Data

Western blot - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

Samples were separated by 7.5% SDS-PAGE.

All lanes:

Western blot - Anti-Mre11 antibody [12D7] - BSA and Azide free (ab214) at 1/1000 dilution

Lane 1:

U87-MG whole cell extracts at 30 µg

Lane 2:

SK-N-SH whole cell extracts at 30 µg

Lane 3:

IMR32 whole cell extracts at 30 µg

Lane 4:

SK-N-AS whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-mouse IgG antibody

false

• WB

Supplier Data

Western blot - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

This image was generated using the ascites version of the product.

7.5% SDS-PAGE

All lanes:

Western blot - Anti-Mre11 antibody [12D7] - BSA and Azide free (ab214) at 1/1000 dilution

Lane 1:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg

Lane 2:

Human Mre-11-transfected HEK-293T whole cell lysate at 30 µg

Secondary

All lanes:

anti-mouse IgG HRP-conjugated antibody

Predicted band size: 80 kDa

false

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

Immunocytochemistry-immunofluorescence using Anti-Mre11 antibody [12D7] - BSA and Azide free, ab214. Publication image from Bohr, V. A. et al., 2016, Nat Commun, 27922005. Legend direct from paper.

WRN inhibits the recruitment of MRE11 and CtIP to DSBs.(a) Confocal microphotographs showing cell cycle independent recruitment of WRN to laser-induced DSBs in U2OS cells. Graph represents data from >200 cells. gH2AX; gamma H2AX. (b) WRN depletion increases MRE11 recruitment to DSBs in G1 cells. siRNA transfected U2OS cells were microirradiated and immunostained for cyclin A2, WRN and MRE11. Graph indicates MRE11 recruitment to laser-induced DSBs in control and WRN siRNA transfected S/G2 and G1 cells. n; number of cells. (c) WRN knockdown promotes CtIP recruitment to laser-induced DSBs. Control (siC) and WRN knockdown U2OS cells were microirradiated to generate DSBs and immunostained with WRN, CtIP and cyclin A2 antibodies. Cyclin A2 positive cells indicate cells in S/G2 phase. (d) Quantitation of CtIP and WRN signals at DSBs as seen in panel E. n=26 cells per set (e) Recruitment kinetics of mCherry-WRN and YFP-MRE11 to laser-induced DSBs in U2OS cells. n=14 cells (f) Real-time recruitment of YFP-MRE11 to laser-induced DSBs in WS cells, AG11395, expressing no WRN (vector control) and mCherry-WRN. n=28 cells (g) Recruitment of GFP-CtIP to laser-induced DSBs in AG11395 WS cells transfected with or without mCherry WRN. n=22 cells. Yellow arrow heads in panel A, B and C indicate laser-induced DSB tracks. a.u., arbitrary unit.

• Other

CiteAb

Other - Anti-Mre11 antibody [12D7] - BSA and Azide free (AB214)

Proximity Ligation Assay using Anti-Mre11 antibody [12D7] - BSA and Azide free, ab214. Publication image from Nieminuszczy, J. et al., 2019, Mol Cell, 31255466. Legend direct from paper.

EXD2 Is Recruited to Stressed Replication Forks(A) Western blot of iPOND samples. Thymidine chase analysis illustrates that EXD2 specifically associates with the replisome. PCNA acts as a control.(B) Schematic of the proximity ligation assay (PLA) employed to detect colocalization of target proteins with nascent DNA.(C) Percentage of cells with MRE11/biotin PLA foci (mean ± SEM, n = 3 independent experiments, t test). Right : representative images of PLA foci (red), DAPI acts as a nuclear counterstain. Scale bar, 10 µm.(D) Percentage of cells with GFP/biotin PLA foci (mean ± SEM, n = 3 independent experiments, t test) in U2OS control cells and U2OS cells expressing GFP-EXD2. Right : representative images of PLA foci (red), DAPI acts as a nuclear counterstain. Scale bar, 10 µm.(E) Laser microirradiation induces rapid redistribution of GFP-EXD2 to damaged chromatin; representative images showing GFP-EXD2 accumulation at laser-generated DNA lesions. GFP-CtIP was used as a positive control. Scale bar, 10 µm.(F) Quantification of GFP-EXD2 (left panel) and GFP-CtIP (right panel) recruitment kinetics (intensity versus time) to laser-generated DNA lesions (mean ± SE, n ≥ 10 cells from 2 independent experiments).