All lanes : Anti-MSH2 antibody [3A2B8C] (ab52266) at 1/2000 dilution

Lane 1 : Cell lysates prepared from human Hela cells
Lane 2 : Cell lysates prepared from A549 cells
Lane 3 : Cell lysates prepared from human A431 cells
Lane 4 : Cell lysates prepared from HEK293 cells

Lysates/proteins at 100 µg per lane.

Secondary
All lanes : HRP-conjugated Goat polyclonal to mouse IgG


Predicted band size: 105 kDa

ab52266 (1/200) detecting MSH2 in HeLa cells (green). Cells were fixed in methanol (-20'C, 10min) and counterstained with DAPI in order to highlight the nucleus. Please refer to abreview for further experimental details.

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ab52266 at 1/1000 dilution staining MSH2 in human Hela cells by Immunocytochemistry/ Immunofluorescence. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG1 was used as secondary antibody. The primary antibody shows green staining in image whilst actin filaments were stained red with Alexa Fluor^® 555 phalloidin.

Overlay histogram showing HeLa cells stained with ab52266 (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 (ab52266, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.

MSH2 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to MSH2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52266.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/10,000 dilution.
Band: 105kDa; MSH2

Immunohistochemical analysis of paraffin-embedded human rectum carcinoma tissue, showing nuclear and cytoplasmic localisation, using ab52266 at a dilution of 1/200 - 1/1000 with DAB staining.