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Product details

Synonyms = mucin 2, oligomeric mucus/gel-forming , MLP , MUC-2 , SMUC

Antibody type = Recombinant Rabbit monoclonal / IgG

Clone = HMV-310

Positive control = Small intestine: A strong MUC2 staining should be seen in goblet cells while other epithelial cells as well as inflammatory cells must remain negative.

Negative control = Small intestine: MUC2 staining must be absent in all non-goblet cells (non-goblet epithelial cells, stroma cells, inflammatory cells, muscular wall).

Cellular localization = Secreted

Reactivity = Human

 

 

Application = Immunohistochemistry
Dilution = 1:100 – 1:200
Intended Use = Research Use Only

Relevance of Antibody

MUC2 is a main protein of intestinal mucus barrier.

Biology Behind

Mucin 2 (MUC2) is an oligomeric mucus gel-forming protein coded by the MUC2 gene at chromosome 11p15.5. The protein is secreted by intestinal goblet cells to the gut where it constitutes the main component of colorectal mucus. The mucins function as a defense mechanism to maintain the integrity of the epithelial cells which are continuously exposed to luminal contents that include large quantities of different bacteria, proteases, bile, and ingested toxins. The mucin barrier consists of two layers. The inner layer is directly attached to the epithelium, densely packed, largely consists of uncleaved MUC2, and is free from bacterial colonization. The outer layer contains bacteria and is less dense due of proteolytic cleavage of MUC2. Under normal conditions, the protease-resistant mucus layer prevents intestinal bacteria to make physical contact with epithelial cells by virtue of its only minute pore sizes and strong hydrophobic properties capable to repel bacteria in the aqueous lumen. Loss of MUC2 confers a microenvironment in which bacteria can activate an inflammatory response at the epithelial surface. The defensive function of MUC2 and of other mucins to maintain the integrity of the epithelial cells can also be exploited by tumor cells in limiting the activation of inflammatory responses. 

Staining Pattern in Normal Tissues

Images describing the MUC2 staining pattern in normal tissues obtained by the antibody HMV-310 are shown in our “Normal Tissue Gallery”.

Brain Cerebrum Negative.
Cerebellum Negative.
Endocrine Tissues Thyroid Negative.
Parathyroid Negative.
Adrenal gland Negative.
Pituitary gland Negative.
Respiratory system Respiratory epithelium Negative.
Lung Negative.
Gastrointestinal Tract Salivary glands Negative.
Esophagus Negative.
Stomach Negative.
Colon Strong MUC2 positivity of mucins in goblet cells.
Duodenum Strong MUC2 positivity of mucins in goblet cells.
Rectum Strong MUC2 positivity of mucins in goblet cells.
Small intestine Strong MUC2 positivity of mucins in goblet cells.
Liver Negative.
Gallbladder Negative.
Pancreas Negative.
Genitourinary Kidney Negative.
Urothelium Negative.
Male genital Prostate Negative.
Seminal vesicles Negative.
Testis Negative.
Epididymis Negative.
Female genital Breast Negative.
Uterus, myometrium Negative.
Uterus, ectocervix Negative.
Uterus endocervix Negative.
Uterus, endometrium Negative.
Fallopian Tube Negative.
Ovary Negative.
Placenta early Negative.
Placenta mature Negative.
Amnion Negative.
Chorion Negative.
Skin Epidermis Negative.
Sebaceous glands Negative.
Muscle/connective tissue Heart muscle Negative.
Skeletal muscle Negative.
Smooth muscle Negative.
Vessel walls Negative.
Fat Negative.
Stroma Negative.
Endothelium Negative.
Bone marrow/lymphoid Bone marrow Negative.
Lymph node Negative.
Spleen Negative.
Thymus Negative.
Tonsil Negative.
Remarks

The findings described above are thus consistent with the RNA data described in the Human Protein Atlas (Tissue expression MUC2). MUC2 staining by HMV-310 was only seen in the small intestine, appendix and the colorectum, which are the only organs for which RNA expression of MUC2 has been documented.

 

Positive control = Small intestine: A strong MUC2 staining should be seen in goblet cells while other epithelial cells as well as inflammatory cells must remain negative.

Negative control = Small intestine: MUC2 staining must be absent in all non-goblet cells (non-goblet epithelial cells, stroma cells, inflammatory cells, muscular wall).

 

Normal tissue gallery

Staining Pattern in Relevant Tumor Types

MUC2 is preferably expressed in a fraction of colorectal adenocarcinomas: It also occurs in breast and stomach cancers as well as (less commonly) in many other tumor entities (preferably adenocarcinoma). 

The TCGA findings on MUC2 RNA expression in different tumor categories have been summarized in the Human Protein Atlas.

 

 

Cancer tissue gallery

Compatibility of Antibodies

No data available at the moment

Protocol Recommendations

IHC users have different preferences on how the stains should look like. Some prefer high staining intensity of the target stain and even accept some background. Others favor absolute specificity and lighter target stains. Factors that invariably lead to more intense staining include higher concentration of the antibody and visualization tools, longer incubation time, higher temperature during incubation, higher temperature and longer duration of the heat induced epitope retrieval (slide pretreatment). The impact of the pH during slide pretreatment has variable effects and depends on the antibody and the target protein.

 

All images and data shown here and in our image galleries are obtained by the manual protocol described below. Other protocols resulting in equivalent staining are described as well.

 

Manual protocol

Freshly cut sections should be used (less than 10 days between cutting and staining). Heat-induced antigen retrieval for 5 minutes in an autoclave at 121°C in pH 7,8 Target Retrieval Solution buffer. Apply HMV-310 at a dilution of 1:150 at 37°C for 60 minutes. Visualization of bound antibody by the EnVision Kit (Dako, Agilent) according to the manufacturer’s directions.

Potential Research Applications

  • The prognostic relevance of MUC2 expression in tumors is not fully clarified.
  • The potential predictive role of MUC2 expression measurement in tumors needs to be further investigated.

Evidence for Antibody Specificity in IHC

There are two ways how the specificity of antibodies can be documented for immunohistochemistry on formalin fixed tissues. These are: 1. Comparison with a second independent method for target expression measurement across a large number of different tissue types (orthogonal strategy), and 2. Comparison with one or several independent antibodies for the same target and showing that all positive staining results are also seen with other antibodies for the same target (independent antibody strategy). 

 

Orthogonal validation: For the antibody HMV-310 specificity is suggested by the complete concordance of the immunostaining data with data from three independent RNA screening studies, including the Human Protein Atlas (HPA) RNA-seq tissue dataset, the FANTOM5 project, and the Genotype-Tissue Expression (GTEx) project, which are all summarized in the human protein atlas Human Protein Atlas (Tissue expression MUC2). MUC2 positivity by HMV-310 is only detectable in the tissues with documented MUC2 RNA expression (colorectum, small intestine, duodenum, appendix). 

 

Comparison of antibodies: True expression of MUC2 in all cell types found MUC2 positive by HMV-310 is corroborated by identical stainings obtained by another commercially available independent antibody (termed “validation antibody”). 





 

 

Normal tissue gallery