MLH1 Mouse Monoclonal Antibody [A9F7]

☑ Relative expression (RE)

Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HCT 116 cell lysate (negative) (15 µg/Lane)
Lane 3: A549 cell lysate (15 µg/Lane)
Lane 4: HepG2 cell lysate (15 µg/Lane)
Lane 5: Jurkat cell lysate (15 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: Lane 1-5 (left): 24 seconds; Lane 1-5 (right): 2 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601135) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: F9 cell lysate (20 µg/Lane)
Lane 3: RAW264.7 cell lysate (20 µg/Lane)
Lane 4: C6 cell lysate (20 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 3 minutes 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601135) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.

☑ Relative expression (RE)

Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: Daudi cell lysate (20 µg/Lane)
Lane 4: K-562 cell lysate (20 µg/Lane)
Lane 5: HL-60 cell lysate (20 µg/Lane)
Lane 6: SW480 cell lysate (20 µg/Lane)
Lane 7: HCT 116 cell lysate (Negative) (20 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 20 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601135) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/2,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si MLH1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 50 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

HA601135 was shown to specifically react with MLH1 in Hela-si NT cells. Weakened band was observed when Hela-si MLH1 sample was tested. Hela-si NT and Hela-si MLH1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (HA601135, 1/2,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-mouse IgG-HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601135) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601135) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601135) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.