Anti-MSH6 antibody [SP93]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [SP93] (AB99889)

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling MSH6 with ab99889 at a dilution of 1/400. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab99889 anti-MSH6 antibody [SP93] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MSH6 antibody [SP93] (AB99889)

Flow cytometry analysis of Hela (human cervix adenocarcinoma) labeling MSH6 with purified ab99889 at 1/20 dilution (5.05 μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabelled cells (blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [SP93] (AB99889)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling MSH6 with ab99889 at 1/100 dilution (1.0 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Sporadically nuclear staining on human colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab99889 for 30 mins at room temperature.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [SP93] (AB99889)

ab99889 staining MSH6 in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab99889 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [SP93] (AB99889)

Staining of MSH6 in a formalin fixed, paraffin embedded Human rectal carcinoma tissue using ab99889 at a dilution of 1/100.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [SP93] (AB99889)

ab99889 staining MSH6 in wild-type HAP1 cells (top panel) and MSH6 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab99889 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [SP93] (AB99889)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue sections labeling MSH6 with ab99889 at 1/100 dilution (1.0 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Nuclear staining on human colon carcinoma, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab99889 for 30 mins at room temperature.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [SP93] (AB99889)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon
tissue sections labeling MSH6 with ab99889 at 1/100 dilution (1.0 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Sporadically nuclear staining on mouse colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab99889 for 30 mins at room temperature.

• WB

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Western blot - Anti-MSH6 antibody [SP93] (AB99889)

ab99889 used in the image is the undiluted version of ab99890.

All lanes:

Western blot - Anti-MSH6 antibody [SP93] (ab99889) at 1/100 dilution

All lanes:

Lysate prepared from A431 cells

Predicted band size: 153 kDa

Observed band size: 170 kDa

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• WB

Supplier Data

Western blot - Anti-MSH6 antibody [SP93] (AB99889)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : MSH6 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : A431 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab99889 observed at 160 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab99889 was shown to specifically react with MSH6 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed in MSH6 knockout samples. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. ab99889 and ab18058 (loading control to Vinculin) were diluted at 1 μg/ml and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-MSH6 antibody [SP93] (ab99889)

Predicted band size: 153 kDa

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