MLH1 Recombinant Mouse Monoclonal Antibody [PSH20-67]
cat.: HA610386
Product Type: Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: PSH20-67
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 85 kDa
Isotype: IgG1
Positive control: SW480 cell lysate, HeLa cell lysate, human appendix tissue, human colon carcinoma tissue, SW480.
Subcellular location: Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell

1:2,000
1:1,000
1:500
Uniprot #: SwissProt: P40692 Human
Alternative names: COCA 2 COCA2 DNA mismatch repair protein Mlh1 FCC 2 FCC2 hMLH 1 hMLH1 HNPCC 2 HNPCC HNPCC2 MGC5172 MLH 1 MLH1 MLH1_HUMAN MutL homolog 1 (E. coli) MutL homolog 1 MutL homolog 1 colon cancer nonpolyposis type 2 MutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) MutL protein homolog 1 MutL, E. coli, homolog of, 1
Images
HA610386_1.jpg Fig1: Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA610386) at 1/2,000 dilution.

Lane 1: SW480 cell lysate (10 µg/Lane)
Lane 2: HCT 116 cell lysate (negative) (10 µg/Lane)
Lane 3: HeLa cell lysate (10 µg/Lane)

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

Exposure time: 1 minute 40 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 (HA610386) at 1/2,000 dilution was used in primary antibody dilution (K1803) 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.
HA610386_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human appendix tissue with Mouse anti-MLH1 antibody (HA610386) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) (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 (HA610386) 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.
HA610386_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-MLH1 antibody (HA610386) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) (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 (HA610386) 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.
HA610386_4.jpg Fig4: Immunocytochemistry analysis of SW480 cells labeling MLH1 with Mouse anti-MLH1 antibody (HA610386) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MLH1 antibody (HA610386) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Negative control : HCT 116 (PMID: 27618981).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.