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☑ 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;
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.
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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;
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.
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☑ 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.
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☑ 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;
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.
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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) 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.
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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) 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.
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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) 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.
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