Unpurified ab92312 staining MLH1 in human colorectal (top) and gastric tissue (bottom) by immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

All lanes : Anti-MLH1 antibody [EPR3894] (ab92312) at 1/10000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MLH1 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 84 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?

This data was developed using the same antibody clone in a different buffer formulation (ab92312).

Lanes 1-4: Merged signal (red and green). Green - ab92312 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab92312 Anti-MLH1 antibody [EPR3894] was shown to specifically react with MLH1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab267223 (knockout cell lysate ab257172) was used. Wild-type and MLH1 knockout samples were subjected to SDS-PAGE. ab92312 and Anti-GAPDH antibody [6C5] - Loading Control were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

 

 

Immunocytochemistry/ Immunofluorescence analysis of SW480 (Human colorectal adenocarcinoma epithelial cell) cells labeling MLH1 with purified ab92312 at 1:500 dilution (1.6 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 anti-alpha tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling MLH1 with purified ab92312 at 1:250 dilution (2.9 µg/ml). Heat mediated antigen retrieval was performed using citrate (pH 6.0) ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling MLH1 with purified ab92312 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control: PBS only

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

Unpurified ab92312 at 1/100 dilution staining MLH1 in human colonic adenocarcinoma by immunohistochemistry, paraffin-embedded tissue. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody.

Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

Unpurified ab92312 (1/200) staining MLH1 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to abreview.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

Overlay histogram showing HeLa cells stained with unpurified ab92312 (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 (ab92312, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).

Unpurified ab92312 at 1/100 dilution staining MLH1 in human tonsil by immunohistochemistry, paraffin-embedded tissue. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody.

Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92312).