MLH-1 antibody (mAb)

Application Notes

Applications Validated by Active Motif:
WB: 2 - 3 µg/ml dilution

For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all blocking solutions may also reduce background. Individual optimization may be required.

Immunogen

This MLH-1 antibody was raised against a synthetic peptide corresponding to human MLH-1.

Buffer

PBS containing 0.02% sodium azide. Sodium azide is highly toxic.

Background

MLH-1 (COCA2 or HNPCC) heterodimerizes with PMS2 to form MutL alpha, a component of the post-replicative DNA mismatch repair system (MMR). DNA repair is initiated by MutS alpha (MSH2-MSH6) or MutS beta (MSH2-MSH6) binding to a dsDNA mismatch, then MutL alpha is recruited to the heteroduplex. Assembly of the MutL-MutS-heteroduplex ternary complex in the presence of RFC and PCNA is sufficient to activate endonuclease activity of PMS2. MLH-1 (COCA2 or HNPCC) introduces single-strand breaks near the mismatch and thus generates new entry points for the exonuclease EXO1 to degrade the strand containing the mismatch. DNA methylation would prevent cleavage and therefore ensure that only the newly mutated DNA strand will be corrected. MutL alpha (MLH1-PMS2) interacts physically with the clamp loader subunits of DNA polymerase III, suggesting that it may play a role to recruit the DNA polymerase III to the site of the MMR. MLH-1 is implicated in DNA damage signaling, a process which induces cell cycle arrest and can lead to apoptosis in the case of major DNA damages. MLH-1 heterodimerizes with MLH-3 to form MutL gamma which plays a role in meiosis.

Positive Control

Whole-cell extracts from Molt-4 cells can be used as positive control for MLH-1 antibody.

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

• ChIP = Chromatin Immunoprecipitation;
• ChIP = ChIP Sequencing;
• CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
• CUT&Tag = Cleavage Under Targets and Tagmentation;
• DB = Dot Blot;
• ELISA = Enzyme-linked Immunosorbent Assay;
• EMSA = Electrophoretic Mobility Shift Assay
• FC = Flow Cytometry;
• ICC = Immunocytochemistry;
• IF = Immunofluorescence;
• IHC = Immunohistochemistry;
• IP = Immunoprecipitation;
• MeDIP = Methyl-DNA Immunoprecipitation;
• NEU = Neutralizing;
• WB = Western Blotting