MLH1 Mouse Monoclonal Antibody [A9F7]
Catalog# HA601135
MLH1 Mouse Monoclonal Antibody [A9F7]
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WB
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IHC-P
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
MLH1 Mouse Monoclonal Antibody [A9F7]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within human MLH1 aa 452-751 / 751.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P
Molecular Weight
Predicted band size: 85 kDa
Positive Control
HeLa cell lysate, A549 cell lysate, HepG2 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, F9 cell lysate, RAW264.7 cell lysate, C6 cell lysate, A431 cell lysate, Daudi cell lysate, K-562 cell lysate, HL-60 cell lysate, SW480 cell lysate, human colon carcinoma tissue, mouse colon tissue, rat small intestine tissue.
Conjugation
unconjugated
Clone Number
A9F7
RRID
Reactivity Data
| WB | IHC-P | |
|---|---|---|
| Human |
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| Mouse |
|
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| Rat |
|
|
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG2b
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IHC-P
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1:1,000
Target
Function
DNA mismatch repair protein Mlh1 or MutL protein homolog 1 is a protein that in humans is encoded by the MLH1 gene located on chromosome 3. It is a gene commonly associated with hereditary nonpolyposis colorectal cancer. Orthologs of human MLH1 have also been studied in other organisms including mouse and the budding yeast Saccharomyces cerevisiae. This gene was identified as a locus frequently mutated in hereditary nonpolyposis colon cancer. It is a human homolog of the E. coli DNA mismatch repair gene, mutL, which mediates protein-protein interactions during mismatch recognition, strand discrimination, and strand removal. Defects in MLH1 are associated with the microsatellite instability observed in hereditary nonpolyposis colon cancer. Alternatively spliced transcript variants encoding different isoforms have been described, but their full-length natures have not been determined.
Background References
1. Guan J et al. MLH1 Deficiency-Triggered DNA Hyperexcision by Exonuclease 1 Activates the cGAS-STING Pathway. Cancer Cell. 2021 Jan
2. Cannavo E et al. Regulation of the MLH1-MLH3 endonuclease in meiosis. Nature. 2020 Oct
Subcellular Location
Nucleus, Chromosome.
Synonyms
COCA 2 antibody
COCA2 antibody
DNA mismatch repair protein Mlh1 antibody
FCC 2 antibody
FCC2 antibody
hMLH 1 antibody
hMLH1 antibody
HNPCC 2 antibody
HNPCC antibody
HNPCC2 antibody
ExpandImages
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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; 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"