Anti-MSH2抗体[3A2B8C]

• Flow Cyt

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Flow Cytometry - Anti-MSH2 antibody [3A2B8C] (AB52266)

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 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH2 antibody [3A2B8C] (AB52266)

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [3A2B8C] (AB52266)

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [3A2B8C] (AB52266)

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.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

• IP

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Immunoprecipitation - Anti-MSH2 antibody [3A2B8C] (AB52266)

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

All lanes:

Immunoprecipitation - Anti-MSH2 antibody [3A2B8C] (ab52266)

Predicted band size: 105 kDa

false

• WB

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Western blot - Anti-MSH2 antibody [3A2B8C] (AB52266)

All lanes:

Western blot - Anti-MSH2 antibody [3A2B8C] (ab52266) at 1/2000 dilution

Lane 1:

Cell lysates prepared from human Hela cells at 100 µg

Lane 2:

Cell lysates prepared from A549 cells at 100 µg

Lane 3:

Cell lysates prepared from human A431 cells at 100 µg

Lane 4:

Cell lysates prepared from HEK293 cells at 100 µg

Secondary

All lanes:

HRP-conjugated Goat polyclonal to mouse IgG

Predicted band size: 105 kDa

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