PRODUCT CODE: ET1604-41

Recombinant MLH1 Monoclonal Antibody (ET1604-41)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of MLH1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1604-41, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Daudi cell lysate<br />
 Lane 2: K562 cell lysate<br />
 Lane 1: A431 cell lysate<br />
 Lane 2: HL-60 cell lysate
  • Western blot analysis of MLH1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1604-41, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Daudi cell lysate<br />
 Lane 2: K562 cell lysate<br />
 Lane 1: A431 cell lysate<br />
 Lane 2: HL-60 cell lysate
  • ICC staining of MLH1 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-41, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat large intestine tissue using anti-MLH1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-41, 1/200) for 30 minutes 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 human tonsil tissue using anti-MLH1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-41, 1/200) for 30 minutes 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 testis tissue using anti-MLH1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-41, 1/200) for 30 minutes 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.
  • Flow cytometric analysis of MLH1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1604-41, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of MLH1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1604-41, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Daudi cell lysate
Lane 2: K562 cell lysate
Lane 1: A431 cell lysate
Lane 2: HL-60 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant MLH1 Monoclonal Antibody (ET1604-41)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Daudi cell lysate, K562 cell lysate, A431 cell lysate, HL-60 cell lysate, HepG2, rat large intestine tissue, human tonsil tissue, mouse testis tissue, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SP08-04

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

85 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:100

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

MLH1

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; MGC5172 antibody; MLH 1 antibody; MLH1 antibody; MLH1_HUMAN antibody; MutL homolog 1 (E. coli) antibody; MutL homolog 1 antibody; MutL homolog 1 colon cancer nonpolyposis type 2 antibody; MutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) antibody; MutL protein homolog 1 antibody; MutL, E. coli, homolog of, 1 antibody

SEQUENCE SIMILARITIES

Belongs to the DNA mismatch repair MutL/HexB family.

TISSUE SPECIFICITY

Colon, lymphocytes, breast, lung, spleen, testis, prostate, thyroid, gall bladder and heart.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

DNA-mismatch repair (MMR) is an essential process in maintaining genetic stability. Lack of a functional DNA-mismatch repair pathway is a common characteristic of several different types of human cancers, either due to an MMR gene mutation or promoter methylation gene silencing. MLH1 is an integral part of the protein complex responsible for mismatch repair and is expressed in lymphocytes, heart, colon, breast, lung, spleen, testis, prostate, thyroid and gall bladder tissues, and is methylated in several ovarian tumors. Loss of MLH1 protein expression is associated with a mutated phenotype, microsatellite instability and a predisposition to cancer. In hereditary nonpolyposis colorectal cancer (HNPCC), an autosomal dominant inherited cancer syndrome that signifies a high risk of colorectal and various other types of cancer, the MLH1 gene exhibits a pathogenic mutation. Certain cancer cell lines, including leukemia CCRF-CEM, colon HCT 116 and KM12, and ovarian cancers SK-OV-3 and IGROV-1, show complete deficiency of MLH1, while MLH1 is expressed in 60% of melanomas, 70% of noninvasive squamous cell carcinomas and 30% of invasive squamous cell carcinomas.