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MLH-1 Recombinant Rabbit Monoclonal Antibody (ZR347), RAbMono™
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Product Details
Z2656RP
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Human
Host/Isotype
Rabbit
/ IgG
Expression System
CHO cells
Class
Recombinant Monoclonal
Type
Antibody
Clone
ZR347
Immunogen
Recombinant full-length of human MLH1 protein
Conjugate
Form
Liquid
Purification
Protein A
Storage buffer
tris with BSA, NP-40
Contains
<0.1% sodium azide
Storage conditions
4°C
Shipping conditions
Ambient (domestic); Wet ice (international)
Product Specific Information
This product is diluted and in a ready-to-use formulation.
A recommended positive control tissue for this product is Colon adenocarcinoma lynch/MLH-1, however positive controls are not limited to this tissue type.
The primary antibody is intended for laboratory professional use in the detection of the corresponding protein in formalin-fixed, paraffin-embedded tissue stained in manual qualitative immunohistochemistry (IHC) testing. This antibody is intended to be used after the primary diagnosis of tumor has been made by conventional histopathology using non-immunological histochemical stains.
This MAb recognizes a protein of 83 kDa, identified as MLH1. Defects in MLH1 are the cause of hereditary non-polyposis colorectal cancer type 2 (HNPCC2). 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 presence of RFC and PCNA is sufficient to activate endonuclease activity of PMS2. It 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 assure that only the newly mutated DNA strand is going to 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. Also implicated in DNA damage signaling, a process, which induces cell cycle arrest and can lead to apoptosis in case of major DNA damages. Heterodimerizes with MLH3 to form MutL gamma, which plays a role in meiosis.
Antibody is used with formalin-fixed and paraffin-embedded sections. Pretreatment of deparaffinized tissue with heat-induced epitope retrieval or enzymatic retrieval is recommended. In general, immunohistochemical (IHC) staining techniques allow for the visualization of antigens via the sequential application of a specific antibody to the antigen (primary antibody), a secondary antibody to the primary antibody (link antibody), an enzyme complex and a chromogenic substrate with interposed washing steps. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with a particular antigen.
A positive tissue control must be run with every staining procedure performed. This tissue may contain both positive and negative staining cells or tissue components and serve as both the positive and negative control tissue. External Positive control materials should be fresh autopsy/biopsy/surgical specimens fixed, processed and embedded as soon as possible in the same manner as the patient sample (s). Positive tissue controls are indicative of correctly prepared tissues and proper staining methods. The tissues used for the external positive control materials should be selected from the patient specimens with well-characterized low levels of the positive target activity that gives weak positive staining. The low level of positivity for external positive controls is designed to ensure detection of subtle changes in the primary antibody sensitivity from instability or problems with the staining methodology. A tissue with weak positive staining is more suitable for optimal quality control and for detecting minor levels of reagent degradation.
Internal or external negative control tissue may be used depending on the guidelines and policies that govern the organization to which the end user belongs to. The variety of cell types present in many tissue sections offers internal negative control sites, but this should be verified by the user. The components that do not stain should demonstrate the absence of specific staining, and provide an indication of non-specific background staining. If specific staining occurs in the negative tissue control sites, results with the patient specimens must be considered invalid.
Target Information
MLH1 is a DNA mismatch repair protein. The repair of mismatch DNA is essential to maintaining the integrity of genetic information over time. An alteration of microsatellite repeats is the result of slippage owing to strand misalignment during DNA replication and is referred to as microsatellite instability (MSI). These defects in DNA repair pathways have been related to human carcinogenesis. The importance of mismatch repair genes became apparent with the identification of the genetic basis for hereditary nonpolyposis colon cancer (HNPC). MSHS2 is involved in the initial cognition of mismatch nucleotides during the replication mismatch repair process. It is thought that after MSH2 binds to a mismatched DNA duplex it is joined by a heterodimer of MLH1 and PMSH, which together help facilitate the later steps in mismatch repair.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: DNA mismatch repair protein Mlh1; DNA mismatch repair protein MutL homolog; human homolog of E. coli mutL gene product, Swiss-Prot Accession Number P23367; MGC5172; MLH-1; mutL homolog 1, colon cancer, nonpolyposis type 2; MutL protein homolog 1; unnamed protein product
Gene Aliases: COCA2; FCC2; hMLH1; HNPCC; HNPCC2; LYNCH2; MLH-1; MLH1; MMRCS1
UniProt ID: (Human) P40692
Entrez Gene ID: (Human) 4292
nuclear-transcribed mRNA poly(A) tail shortening
resolution of meiotic recombination intermediates
somatic recombination of immunoglobulin genes involved in immune response
DNA repair
mismatch repair
double-strand break repair via nonhomologous end joining
response to stress
cellular response to DNA damage stimulus
male meiosis chromosome segregation
synapsis
reciprocal meiotic recombination
male meiosis
spermatogenesis
intrinsic apoptotic signaling pathway in response to DNA damage
response to bacterium
female meiosis chromosome segregation
somatic hypermutation of immunoglobulin genes
somatic recombination of immunoglobulin gene segments
meiotic metaphase I plate congression
meiotic chromosome segregation
meiotic telomere clustering
homologous chromosome segregation
isotype switching
negative regulation of mitotic recombination
positive regulation of isotype switching to IgA isotypes
positive regulation of isotype switching to IgG isotypes
oogenesis
meiotic spindle midzone assembly
meiotic cell cycle
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Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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